Am Ind Hyg Assoc J, 1987 Aug, 48(8), 745 - 51 A study of the relationship between airborne contaminants and environmental factors in Dutch swine confinement buildings; Attwood P et al.; A total of 171 swine confinement buildings were studied to determine the concentrations of airborne total and D50 less than or equal to micron 8.5 dust fractions, total and gram-negative bacteria, bacterial endotoxin and NH3 . The concentrations of these airborne contaminants then were correlated statistically to a number of environmental factors such as feeding practices, number of animals and ventilation parameters . The results showed that airborne dust, endotoxin, bacteria and NH3 are commonly in high concentrations within the swine confinement buildings and that these are at levels where health effects have been observed in other studies . Correlation of these airborne contaminants to a number of environmental factors showed that while ventilation is an important criteria for airborne contaminants, there are a number of farming practices that significantly contribute to the levels of airborne contaminants currently found . Pearson correlations indicate a number of important criteria that the industrial hygienist should measure when faced with problems in agricultural confinement buildings. Forensic Sci Int, 1987 Aug, 34(4), 255 - 6 Death caused by DF-2, following a dog-bite; Theilade P et al.; The Gram-negative rod DF-2 was isolated from blood-cultures, the blood sample taken 3 days after death from a 65-year-old woman who had been bitten in her finger by her dog . At the medico-legal external examination marbling of the skin was found, suggesting septicemia . In persons found dead with a history of dog exposition and with no other obvious cause of death, examination for DF-2 should be performed. J Bacteriol, 1987 Aug, 169(8), 3654 - 63 Nephelometric determination of turgor pressure in growing gram-negative bacteria; Koch AL et al.; Gas vesicles were used as probes to measure turgor pressure in Ancylobacter aquaticus . The externally applied pressure required to collapse the vesicles in turgid cells was compared with that in cells whose turgor had been partially or totally removed by adding an impermeable solute to the external medium . Since gram-negative bacteria do not have rigid cell walls, plasmolysis is not expected to occur in the same way as it does in the cells of higher plants . Bacterial cells shrink considerably before plasmolysis occurs in hyperosmotic media . The increase in pressure required to collapse 50% of the vesicles as external osmotic pressure increases is less than predicted from the degree of osmotically inducible shrinkage seen with this organism or with another gram-negative bacterium . This feature complicates the calculation of the turgor pressure as the difference between the collapse pressure of vesicles with and without sucrose present in the medium . We propose a new model of the relationship between turgor pressure and the cell wall stress in gram-negative bacteria based on the behavior of an ideal elastic container when the pressure differential across its surface is decreased . We developed a new curve-fitting technique for evaluating bacterial turgor pressure measurements. Plast Reconstr Surg, 1987 Aug, 80(2), 213 - 25 Craniofacial infection in 10 years of transcranial surgery; David DJ et al.; Infection following transcranial surgery may be devastating . A review of 170 transcranial operations is presented with a focus on postoperative infection and its relationship to patient age, preoperative microbiology, pattern of operation, length of operation, and the use of antibiotic prophylaxis . The overall postoperative infection rate was 6.5 percent, but the infection rate in adults (23.5 percent) was much higher than in children (2.2 percent) . Higher infection rates were found in adults with craniofacial dysostoses undergoing lengthy frontofacial advancements which required tracheostomy airway management . The residual frontal extradural dead space following advancement in adults is a sanctuary to infecting organisms from the respiratory tract--especially Pseudomonas transferred from the tracheostomy site into the upper airway and intracranial dead space by ventilation forces . Operating times for patients who became infected were 2 1/2 hours longer than average operating times for transcranial operations . Preoperative microbiology of the craniofacial region was not a good predictor of subsequent infection . Recommendations include operative intervention at an early age, short preoperative hospital stay, antibiotic prophylaxis to include gram-negative cover, surgical measures to either fill or isolate the dead space, and strict tracheostomy care--preferably with the patient being barrier-nursed. Eur J Clin Microbiol, 1987 Aug, 6(4), 456 - 9 Emergence of resistance in gram-negative bacteria during therapy with expanded-spectrum cephalosporins; Dworzack DL et al.; To assess the clinical importance of emergence of beta-lactam resistance caused by stable derepression of chromosomal beta-lactamases, sequential cultures from patients treated with expanded-spectrum cephalosporins were monitored for the persistence of bacteria possessing these enzymes . Antibiotic susceptibilities and beta-lactamase production before and after cefoxitin induction were determined in sequential isolates of individual bacterial strains . Of 49 strains isolated from 44 patients, 25 strains (51%) were eradicated by cephalosporin therapy, 17 strains (35%) persisted with unchanged susceptibility in sequential cultures, and 7 strains (14%) from 7 patients developed multiple beta-lactam resistance during cephalosporin therapy . In 6 of the 7 strains, resistance was associated with stable derepression of beta-lactamases . In the patient group whose strains developed resistance, subsequent use of non-beta-lactam antibiotics was more frequent and mortality was higher. Immunology, 1987 Aug, 61(4), 527 - 33 Characterization of an antigen secreted by Chlamydia-infected cell culture; Stuart ES et al.; A soluble genus-specific chlamydial antigen has been isolated from the supernatants of cultures infected with Chlamydia trachomatis and from other sources . The antigen is a glycolipid that is secreted during the infective cycle . This exoglycolipid can be hydrolysed and fractionated into polysaccharide and lipid components . Both fractions retain antigenic activity . An immunodominant antigenic determinant of the lipid component contains fatty acids of C17 and C18:1 . The polysaccharide immunodominant epitope gives rise to gulose when derivatives are formed . The secretion of the antigen into the media supernatant, the presence of gulose and the observed molecular weight are consistent with properties of alginate secreted by Gram-negative bacteria . Chemical analyses and SDS-PAGE indicate that the exoglycolipid is markedly different from LPS. J Clin Pharm Ther, 1987 Aug, 12(4), 249 - 54 A comparison of two incubation temperatures for the isolation of gram-negative contaminants from raw materials and non-sterile pharmaceuticals; Ferguson A et al.; Selective and non-selective broth enrichment techniques may be used in the isolation of microbial contaminants from pharmaceutical products . A non-selective method may give better recovery rates for damaged organisms . A trial was carried out to determine whether the recovery of Gram-negative contaminants could be improved by using an incubation temperature of 30 degrees C for 48 h, rather than the more widely used 37 degrees C for 24 h . Contaminants were isolated from 3.2% of samples incubated at the lower temperature compared with 0.8% at the higher temperature . The recovery rate from raw materials improved noticeably (9.0% compared with 0.9%). Anal Biochem, 1987 Aug 1, 164(2), 320 - 30 A highly efficient procedure for the quantitative formation of intact and viable lysozyme spheroplasts from Escherichia coli; Marvin HJ et al.; This paper describes a highly efficient procedure for the quantitative conversion of Escherichia coli cells to spheroplasts utilizing 100- to 1000-fold less lysozyme than in the most efficient procedures used to date . The resulting spheroplasts have intact outer and inner membranes and are fully viable on agar plates . The spheroplasting procedure is a refinement of earlier procedures and enables regulation of the translocation of minute amounts of lysozyme into the periplasmic space of E . coli cells, based on a Ca2+ pretreatment, an EDTA incubation, and a heat shock . About 1000 lysozyme molecules per cell are sufficient for complete spheroplast formation (greater than 98%) . Some of the characteristics of these spheroplasts prior to and after recovery are described . It is anticipated that such viable spheroplasts will be useful in the study of fusion of gram-negative cells and other membrane systems, in the introduction of DNA and proteins into refractory gram-negative cells, and in investigating envelope-related synthesis and assembly processes. Am J Physiol, 1987 Aug, 253(2 Pt 1), E123 - 9 Altered glucose kinetics in diabetic rats during gram-negative infection; Lang CH et al.; The present study examined the purported exacerbating effect of sepsis on glucose metabolism in diabetes . Diabetes was induced in rats by an intravenous injection of 70 or 45 mg/kg streptozotocin . The higher dose produced "severe" diabetes, whereas the lower dose of streptozotocin produced a milder, "latent" diabetes . After a chronic diabetic state had developed for 4 wk, rats had catheters implanted and sepsis induced by intraperitoneal injections of live Escherichia coli . After 24 h of sepsis the blood glucose concentration was unchanged in nondiabetics and latent diabetics, but glucose decreased from 15 to 8 mM in the septic severe diabetic group . This decrease in blood glucose was not accompanied by alterations in the plasma insulin concentration . Glucose turnover, assessed by the constant intravenous infusion of {6-3H}- and {U-14C}glucose, was elevated in the severe diabetic group, compared with either latent diabetics or nondiabetics . Induction of sepsis produced a slight decrease in the glucose turnover in the severe diabetic group but did not alter turnover in the latent diabetics . The rate of glucose disappearance, used to quantitate the alterations in plasma glucose after an intravenous glucose tolerance test, was decreased in both groups of diabetics and was proportional to the severity of the diabetic state . Sepsis increased the rate of glucose disappearance in nondiabetic rats but had no effect in either group of diabetic animals . Sepsis also failed to alter the insulinogenic index, used to estimate the insulin secretory capacity, in diabetic rats.(ABSTRACT TRUNCATED AT 250 WORDS) Cancer, 1987 Jul 15, 60(2), 255 - 62 Imipenem/cilastatin therapy of infections in cancer patients; Bodey GP et al.; Imipenem/cilastatin was administered during 153 febrile episodes occurring in cancer patients and the response rate was 68% . Considering only documented infections the response rate was 71% . Patients who received imipenem as initial therapy had a higher response rate than patients who received it after failing other antibiotics (77% versus 68%) . The overall response rates for septicemias and pneumonias were 75% and 58% . Among the 57 gram-negative infections 77% responded, but the response rate was substantially higher if imipenem was used as initial therapy (94% versus 69%) . The poorest response rate was observed when imipenem was given as secondary therapy for Pseudomonas infections (50%), but most of these patients had failed to respond to other appropriate antibiotics . The only serious side effect was seizures which occurred in ten patients, although eight of them had other predisposing factors . Imipenem appears to be a useful antibiotic for treatment of infections, even in neutropenic cancer patients. Appl Environ Microbiol, 1987 Jul, 53(7), 1718 - 20 A rapid test for chitinase activity that uses 4-methylumbelliferyl-N-acetyl-beta-D-glucosaminide; O'Brien M et al.; A total of 101 strains of bacteria from environmental and clinical sources, most of which were gram negative, were tested for chitobiase activity by using a filter paper spot test with 4-methylumbelliferyl-N-acetyl-beta-D-glucosaminide as the substrate . The results were compared with those obtained by a conventional plate method for chitinase activity by using colloidal chitin as the substrate . There was excellent agreement in the results for both methods . The filter paper spot test with 4-methylumbelliferyl-N-acetyl-beta-D-glucosaminide has the advantages of being rapid, simple to perform, and inexpensive . This method should be adaptable to a wider range of microorganisms, particularly those with unusual growth requirements. Appl Environ Microbiol, 1987 Jul, 53(7), 1685 - 9 Bacterial detoxification of diisopropyl fluorophosphate; Attaway H et al.; The ability of 18 gram-negative bacterial isolates to detoxify diisopropyl fluorophosphate, a structural analog of the agents soman and sarin, was investigated . Detoxification by both frozen cell sonicates and acetone powders was assayed by two methods, i.e., the hydrolytic release of fluoride, measured by a fluoride-specific ion electrode, and the disappearance of acetylcholinesterase inhibition in vitro . Frozen cell sonicates for all strains exhibited some activity (F- ion release) . In general, acetone powder preparations produced higher activity than frozen cell sonicates did, and the highest activities were exhibited by strains with known parathion hydrolase activity . Two ranges in activity were observed, low level, ranging from 0.1 to 7.0 mumol/min per g of protein, and high level, detected only in parathion hydrolase-producing strains, from 47 to greater than 300 mumol/min per g of protein . Results indicate that parathion hydrolase was nonspecific in phosphoesterase activity . Also, it was an effective detoxicant at low concentrations and near-neutral pH. Thromb Res, 1987 Jul 1, 47(1), 37 - 46 Experimental gram-negative septicemia: thromboplastin generation in mononuclear phagocytes from different anatomical sites; Almdahl SM et al.; Rats were subjected to gram-negative septicemia induced by cecal perforation or were sham-operated . Thromboplastin values increased in blood monocytes (40-fold), peritoneal macrophages (115-fold) pleural macrophages (5-fold), splenic macrophages (3-fold), and lung alveolar macrophages (1.4-fold) in septic animals as compared to controls . In septic animals disseminated intravascular coagulation was evidenced by a significant (p less than 0.05) fall in fibrinogen, factor VII, X and platelets . A simultaneous and significant (p less than 0.05) decrease in thromboplastin content of tissue-specimens from lung and spleen was observed in rats with septicemia, whereas increased thromboplastin values were demonstrated in tissue-samples from cecum - the infectious focus . This might reflect mobilization of mononuclear phagocytes in favour of the site of infection. J Clin Pharmacol, 1987 Jul, 27(7), 491 - 8 The novel therapeutic implications of azlocillin's dose-dependent pharmacokinetics: contributing physiologic mechanisms and a prospective, cross-over designed trial; Whelton A et al.; Azlocillin is an important acylureido penicillin antibiotic for the management of complex gram-negative infections particularly those caused by Pseudomonas species . The current studies demonstrate that it manifests dose-dependent pharmacokinetics during the usual regimens of clinical dosing, that enterohepatic recirculation does not occur and that renal tubular secretion (maximum renal tubular secretory capacity 300 +/- 30 micrograms/min) and hepatic metabolism appear to be the dominant contributors to the dose-dependent nature of azlocillin . The possible therapeutic implications of azlocillin's dose dependency were evaluated by undertaking a six-day randomized, prospective, cross-over design study to evaluate the pharmacokinetic disposition of the drug during a 3-g q4h (typically used in adults) regimen versus a 5-g q8h regimen . By using the area under the serum-time concentration curve (AUC) as the major comparative parameter for these two regimens, the results demonstrate that both regimens provide approximately equal quantitative amounts of the drug systemically as a result of azlocillin's dose dependency . The AUC values, although not therapeutic end points, nonetheless correlate well with clinical response to antibiotic therapy . The 5-g q8h regimen was well tolerated . It is less disruptive for patients, requires half the number of intravenous administrations, 17% less drug, and is more cost effective than the 3-g q4h regimen. Ann Intern Med, 1987 Jul, 107(1), 36 - 41 The multifactorial basis for hypocalcemia during sepsis . Studies of the parathyroid hormone-vitamin D axis; Zaloga GP et al.; To learn about the pathogenesis of sepsis-associated hypocalcemia, we measured serum ionized calcium concentrations in 60 critically ill patients with bacterial sepsis; 12 (20%) had hypocalcemia . The mortality rate in the hypocalcemic patients with sepsis (50%) was higher than that in the normocalcemic patients with sepsis (29%) . Only patients with gram-negative sepsis became hypocalcemic, and hypocalcemia contributed to hypotension in 7 of the 12 hypocalcemic patients . Serum calcium concentrations returned to normal in each of those patients with sepsis who survived . Hypocalcemia during sepsis occurred in previously normocalcemic patients and was multifactorial in origin, resulting from acquired parathyroid gland insufficiency, renal 1 alpha-hydroxylase insufficiency, vitamin D deficiency, and acquired calcitriol resistance . We conclude that the hypocalcemia of sepsis is associated with a high mortality rate and usually occurs in previously normocalcemic patients who acquire a defect in the parathyroid-vitamin D axis. Mol Microbiol, 1987 Jul, 1(1), 29 - 36 Sensitivity of Escherichia coli to various beta-lactams is determined by the interplay of outer membrane permeability and degradation by periplasmic beta-lactamases: a quantitative predictive treatment; Nikaido H et al.; In Gram-negative bacteria, beta-lactam antibiotics must overcome two barriers, the outer membrane and the periplasmic beta-lactamase, before they reach the targets of their action, penicillin-binding proteins . Although the barrier property of the outer membrane and catalytic property of the beta-lactamases have been studied and their significance in creating beta-lactam resistance emphasized, the interaction between these two barriers has not been treated quantitatively . Such treatment shows that the sensitivity, to a variety of beta-lactams, of the Escherichia coli K-12 cells containing very different levels of chromosomally coded AmpC beta-lactamase, or a plasmid-coded TEM-type beta-lactamase, can be predicted rather accurately from the penetration rate through the outer membrane and the hydrolysis rate in the periplasm . We further propose a new parameter, 'target access index', which is a quantitative expression of the result of interaction between the two barriers, and reflects the probability of success for the antibiotic to reach the targets. J Antimicrob Chemother, 1987 Jul, 20(1), 37 - 45 Effect of a diazaborine derivative (Sa 84.474) on the virulence of Escherichia coli; Lam C et al.; The scope of selective inhibition of lipopolysaccharide (LPS) biosynthesis in virulent organisms with a diazaborine derivative (Sa 84.474) as a means to render them avirulent was explored . A serum resistant Escherichia coli 0111:B4 became serum sensitive following its cultivation in vitro in media containing 1.5 mg/l of Sa 84.474, a concentration shown previously to inhibit over 95% of normal LPS biosynthesis in its mutant J5 strain . An encapsulated E . coli 01:K1 was also converted to serum sensitivity . In addition, Sa 84.474-pretreatment increased the efficiency with which the previously resistant organisms were opsonized in normal human serum and subsequently phagocytosed by granulocytes . In vivo, intravenously inoculated, Sa 84.474-pretreated bacteria were rapidly removed from the blood circulation and were significantly (P less than 0.05) less virulent in inducing lethal peritoneal infections in mice (LD50 4.9 +/- 1 X 10(6) cfu) when compared to untreated control bacteria (LD50 2.2 +/- 0.97 X 10(6) cfu) . The results suggest that the LPS plays an important role in the virulence of the two bacterial strains and imply that agents acting in a similar manner but with acceptable selective toxicity might be novel drugs for therapy of Gram-negative bacterial infections. J Infect, 1987 Jul, 15 Suppl 1, 21 - 8 The use of intravenous immunoglobulin for the treatment of infection: an overview; Yap PL; Immunoglobulin replacement therapy in the form of intravenous immunoglobulin (IVIgG) is clearly of benefit in primary hypogammaglobulinaemia and related disorders involving antibody deficiency . However, its use in the prevention of infection in other conditions is controversial and needs to be clarified . IVIgG therapy may be of benefit in selected patients with IgG subclass deficiency, and with a proven history of recurrent upper respiratory tract infections . IVIgG therapy may also benefit infants with AIDS and recurrent bacterial infections, but is only rarely of value in cases of adult AIDS . The use of 'normal' IVIgG from unselected blood donors to treat viral infections, infections due to gram negative organisms, or to neutralise endotoxin, is probably not indicated on theoretical and practical grounds, and because of the high cost . IVIgG preparations derived from plasma donations selected for high specific antibody levels to the relevant microorganism will probably be of greater benefit than normal IVIgG in patients with specific infections. Proc Natl Acad Sci U S A, 1987 Jul, 84(13), 4645 - 9 Transposition of bacteriophage Mu in the Legionnaires disease bacterium; Mintz CS et al.; Legionnaires disease is an acute respiratory disease that is often fatal for immunocompromised patients . The causative agent of this disease, Legionella pneumophila, is a Gram-negative bacterium that is present in a variety of aquatic environments . L . pneumophila is a facultative intracellular parasite; it grows within human phagocytic cells and eventually causes their destruction . In contrast to many other intracellular parasites, L . pneumophila is a Gram-negative bacterium that can be grown in standard microbiological culture medium . To determine the factors that enable this organism to enter, survive, and multiply within human mononuclear phagocytes, we chose bacteriophage Mu, a powerful genetic tool that transposes within the host cell genome, to generate insertion mutations and gene fusions in the Legionella genome . Certain derivatives of Mu are able to generate fusions between target genes and the lac operon from Escherichia coli . We have determined that although Mu is unable to attach to L . pneumophila or complete its life cycle within Legionella, it does transpose within the Legionella genome . Transposition was detected with a mini-Mu phage that carries the lac operon of E . coli. Eur J Biochem, 1987 Jul 1, 166(1), 131 - 7 The soluble c-type cytochromes from the bacterium Aquaspirillum itersonii . The complete amino acid sequence of the cytochrome c-550; Woolley KJ; A complete amino acid sequence is proposed for the cytochrome c-550 isolated from the gram-negative chemo-organotrophic bacterium Aquaspirillum itersonii . The sequence, a single polypeptide chain of 111 residues, was deduced from the sequences of peptides obtained by tryptic, thermolytic or chymotryptic digestion . The cytochrome shows a high degree of sequence homology with the cytochrome c2 from the photosynthetic bacterium Rhodospirillum rubrum, and the evolutionary implications of this are considered. Eur J Biochem, 1987 Jul 1, 166(1), 127 - 30 Purification and properties of the soluble cytochromes c-550 and c-556 from the bacterium Aquaspirillum itersonii; Woolley KJ; Two c-type cytochromes were isolated from cells of the gram-negative bacterium Aquaspirillum itersonii grown under low aeration in the presence of nitrate . The major component, cytochrome c-550, was equated with the (single) c-type cytochrome previously reported to be present in this organism {Clark-Walker, G . D . & Lascelles, J . (1970) Arch . Biochem . Biophys . 136, 153-159}, although a significantly higher molecular mass was apparent in the present work . The complete amino acid sequence of this cytochrome is reported in the accompanying paper . A second soluble c-type cytochrome, designated c-556, was also isolated . The molecular mass, isoelectric point, spectrum, midpoint oxidation reduction potential and amino acid composition of this monoheam cytochrome are reported . The possible relationship of this cytochrome to other cytochromes c-556 is discussed. Jpn J Antibiot, 1987 Jun, 40(6), 1129 - 34 {Fundamental study of amikacin in the newborn}; Hashira S et al.; Amikacin (AMK) is one of the aminoglycoside antibiotics, derived from kanamycin A . It has a broad spectrum against Gram-negative rods but its usefulness is mainly in the efficacy against Gram-negative rods which do not respond to commonly used kanamycin and gentamicin . The efficacy and the safety of AMK have been confirmed in children and mature babies . In the trial reported here, we evaluated AMK in newborn . 1 . AMK was administered to 13 mature and 8 premature babies via intramuscular injection or intravenous drip infusion for 30 minutes or 1 hour and its blood concentrations were determined . These administrations resulted in blood concentrations 4.47-9.67 mcg/ml with dosage levels 2.3 mg/kg (mean 6.92 +/- 1.66 mcg/ml), 5.86-26.1 mcg/ml with 5-6 mg/kg (mean 15.4 +/- 4.63 mcg/ml) and 27.5-37.7 mcg/ml with 7.5 mg/kg (mean 31.0 +/- 4.76 mcg/ml) . Blood half-lives were 1.88 to 9.66 hours, showing longer half-lives in younger subjects . 2 . Exchange transfusion (150-180 ml/kg) was performed in 5 mature babies and the variation of blood concentrations of AMK was studied . The study showed that blood concentrations of AMK after the exchange transfusion were 25.6-41.5% (mean 32.3 +/- 5.4%) of the levels detected before the transfusion. Am Ind Hyg Assoc J, 1987 Jun, 48(6), 511 - 4 Isolation and identification of gram negative bacteria from raw baled cotton and synthetic textile fibers with special reference to environmental GNB and endotoxin concentrations of textile mill; Gokani VN et al.; The objective of this study was to examine the gram-negative bacterial (GNB) content of Indian raw baled cotton fibers and to compare with the U.S . cottons . Airborne endotoxin also was estimated in the different work places of the mill . On comparison with data on U.S . cottons, GNB content was found to be as high as in U.S . cottons . Moreover, endotoxin concentration of cardroom dust from an Indian cotton mill was found to be significantly (P less than 0.01) higher than in the research laboratory of the same cotton mill as well as in the cardroom of a synthetic mill. Lab Anim Sci, 1987 Jun, 37(3), 341 - 4 Epidermal capillariasis in South African clawed frogs (Xenopus laevis); Stephens LC et al.; Significant morbidity and mortality of South African clawed frogs, Xenopus laevis, can be caused by parasitism of the epidermis by a capillarid nematode . These worms produce an erosive dermatitis that is complicated by infection with gram-negative microorganisms . The nematode apparently has a direct life cycle that can be completed within the epidermis of the frog . These characteristics are suggested by the lack of an intermediate host and failure to detect visceral migration . Another unique feature of the parasite was the presence of larvated eggs, in utero . Some have named the parasite Capillaria xenopodis, whereas others called it Pseudocapillaroides xenopi to distinguish it form the typical members of the genus Capillaria. Hepatogastroenterology, 1987 Jun, 34(3), 123 - 6 Circulating lipid A antibodies and their relationship to different clinical conditions of patients with Crohn's disease; Kruis W et al.; Endotoxins have been suggested to be a factor in the pathophysiology of Crohn's disease (CD) . We determined circulating antibodies against lipid A, a component common to endotoxins of gram-negative bacteria . Lipid A antibody titers in 91 patients with CD were significantly higher than in 56 patients with ulcerative colitis and 68 healthy subjects . In active CD lipid A antibodies were higher than in quiescent CD and markedly elevated titers of lipid A antibodies were associated with a severe course of CD . Duration, extent and localisation of CD showed no relationship to antibody titers against lipid A . Patients with prior bowel resection had a tendency towards lower antibody titers in comparison with non-operated patients . After total removal of inflamed bowel tissue, lipid A antibodies frequently disappeared . Medical therapy had different effects: prednisone and sulfasalazine did not influence antibody formation against lipid A, whereas antibody titers dropped significantly after therapy with ampicillin . These results confirm elevated circulating lipid A antibodies in patients with CD . Although it remains unclear whether lipid A antibodies are only an epiphenomenon in CD, on the basis of this clinical study further evidence is provided for the involvement of lipid A in the pathophysiology of CD. J Bacteriol, 1987 Jun, 169(6), 2881 - 4 Putrescine and cadaverine are constituents of peptidoglycan in Veillonella alcalescens and Veillonella parvula; Kamio Y et al.; Veillonella alcalescens ATCC 17745, a strictly anaerobic, gram-negative small coccus, requires putrescine or cadaverine for growth (M . B . Ritchey, and E . A . Delwiche, J . Bacteriol . 124:1213-1219, 1975) . Both putrescine and cadaverine were demonstrated to be incorporated exclusively into the peptidoglycan layer of V . alcalescens ATCC 17745 . V . parvula GAI 0574 also proved to contain putrescine as a component of peptidoglycan . The primary chemical structure of the peptidoglycan common to the two Veillonella species is N-acetylglucosamine-N-acetylmuramic acid-L-alanine-D-glutamic acid gamma-meso-diaminopimelic acid-D-alanine . Putrescine or cadaverine links covalently to the alpha-carboxyl group of the D-glutamic acid residue of the peptidoglycan is necessary for normal cell growth . In V . alcalescens ATCC 17745, above 40% saturation at cadaverine linked to the alpha-carboxyl group of the D-glutamic acid residue of the peptidoglycan is necessary for normal growth. Brain Behav Immun, 1987 Jun, 1(2), 159 - 72 Significance of increased secretion of glucocorticoids in mice and rats injected with bacterial endotoxin; Nakano K et al.; Injection of mice or rats with lipopolysaccharide (LPS) is associated with an increased secretion of glucocorticoids . The high level of mortality following injection of LPS that is noted in adrenalectomized rats can be reversed by dexamethasone or corticosterone . That histamine may be an endogenous mediator of the release of corticosterone caused by LPS is suggested by an attenuation of this corticosterone response by promethazine, an H1 antihistamine . Additional support that LPS-dependent glucocorticoid secretion is mediated, in part, by histamine, is suggested by spleen cell transfer studies revealing differences in the induction of histidine decarboxylase (HDC) synthesis and corticosterone release by the C3H/HeN and C3H/HeJ strains of mice that are differentially sensitive to LPS effects . These and other data on increased levels of histamine and HDC during mitogen-induced lymphocyte blastogenesis, as well as experiments revealing immunomodulatory effects of histamine and histamine agonists and antagonists on lymphocyte blastogenesis, are consistent with the hypothesis that following infection with gram-negative bacteria, the histamine-induced increase in glucocorticoid secretion results in inhibition of HDC in splenocytes, a concomitant attenuation of histamine production, and a resulting return to immune homeostasis. Biochem J, 1987 Jun 1, 244(2), 393 - 9 Bacterial killing by complement . C9-mediated killing in the absence of C5b-8; Dankert JR et al.; The ability of serum complement to kill Gram-negative bacteria requires assembly of the membrane attack complex (MAC) on the cell surface . The molecular events that lead to cell killing after MAC assembly are unknown . We have investigated the effect of C9 on bacterial survival in the presence and absence of its receptor, the C5b-8 complex, on the outer membrane . A fluorescence assay of the membrane potential across the inner bacterial membrane revealed that addition of C9 to cells bearing the performed C5b-8 complex caused a rapid and complete dissipation of the membrane potential . No fluorescence change was observed in serum-resistant strains of Escherichia coli . Addition of trypsin, after C9 was bound to C5b-8, did not rescue the cells from the lethal effects of C9 . Furthermore, assays of cell killing kinetics and C9 binding indicate that formation of tubular poly(C9) is not required for killing . When C9 was introduced into the periplasmic space in the absence of its receptor by means of an osmotic shock procedure, cell killing occurred . Other proteins, such as C8 or serum albumin, were not toxic, and C9 was ineffective against two resistant strains . The results presented here and previously {Dankert & Esser (1986) Biochemistry 25, 1094-1100}, when considered together, indicate that the 'lethal unit' in complement killing of some Gram-negative bacteria is a C9-derived product that acts by dissipation of cellular energy. J Med Virol, 1987 Jun, 22(2), 189 - 98 Microbial structures in a patient with sporadic non-A, non-B fulminant hepatitis treated by liver transplantation; Fagan EA et al.; Double-shelled virus-like particles (60 nm) and long cytoplasmic tubular structures were found in the cytoplasm of hepatocytes from areas of collapsed and regenerating areas of hepatectomised liver in a 13-year-old boy who received a liver graft for fulminant hepatitis attributed to sporadic non-A, non-B hepatitis . The patient died on the ninth postoperative day from acute graft failure . Although virus-like particles were not found, instead, gram-negative rods were identified in the necrotic graft and the most likely cause of death was a gram-negative septicaemia with a Shwartzman-like reaction localized to the liver. Am J Clin Pathol, 1987 Jun, 87(6), 739 - 44 Ultrastructural observations in cat scratch disease; Osborne BM et al.; Because the causative bacterium of cat scratch disease has not been definitively cultured or fully characterized, the authors have studied its ultrastructure in lymph node biopsies from two patients using glutaraldehyde-fixed tissue . In both specimens, the organisms were invariably extracellular, forming small groups within bundles of collagen fibrils . Their appearance was similar in necrotic and viable regions of the nodes, although in the latter sites they could not be identified by light microscopic examination with the Warthin-Starry stain . The bacteria were pleomorphic rods, and, despite faint gram-negative staining, their walls were consistently thick and homogeneous. J Clin Pharmacol, 1987 May-Jun, 27(5), 419 - 24 Predictive performance of Sawchuk-Zaske and Bayesian dosing methods for tobramycin; Rodvold KA et al.; We evaluated the reliability of pharmacokinetic parameter estimations determined by the Sawchuk-Zaske method and by a Bayesian method for predicting steady-state tobramycin concentrations on day 4 and day 10 of therapy in 30 patients treated for gram-negative infections . We also assessed the effect of using only trough tobramycin concentrations on the predictive performance of the Bayesian method . The mean estimation of tobramycin volume of distribution was significantly different for the Bayesian methods compared with the Sawchuk-Zaske methods when the same number of L/kg) . Comparing the Bayesian and Sawchuk-Zaske methods when the same number of serum concentrations was available, the Bayesian method overpredicted peak concentrations on day 4 of therapy . When only trough concentration data were used, the Bayesian method significantly overpredicted peak concentrations compared with the Sawchuk-Zaske method on days 4 and 10 of therapy . Each method predicted steady-state trough concentrations without significant differences in bias or accuracy . The Bayesian method may be useful in providing aminoglycoside dosing recommendations. Zh Mikrobiol Epidemiol Immunobiol, 1987 May, (5), 3 - 9 {The problem of predicting new species of microorganisms (the example of nonfermenting gram-negative microorganisms)}; Kalina GP; Various microbial species, embraced by the genera, have been found to correspond to their phylogeny by the types of nutrition and the levels of pathogenicity . A relationship between the types of nutrition and the levels of parasitism and pathogenicity has been revealed . The results of analysis make it possible to assume, with a great degree of probability, the existence of new species (subspecies, variants) in the environment and/or as agents of diseases of unclear etiology. Antimicrob Agents Chemother, 1987 May, 31(5), 805 - 7 Clinical efficacy and levels of ciprofloxacin in tissue in patients with soft tissue infection; Licitra CM et al.; The clinical efficacy of ciprofloxacin was evaluated with 21 patients with soft tissue infection due mainly to gram-negative aerobic bacteria . Clinical cure was noted in 16 patients (76%), and clinical improvement was noted in the remaining 5 patients (24%) . In addition, levels of ciprofloxacin were measured in the sera and tissues of 11 patients . Mean concentrations in tissue averaged 1.75 times the levels in serum . Ciprofloxacin should provide serum and soft tissue concentrations above the MICs for most gram-negative aerobic bacteria. Pediatr Infect Dis J, 1987 May, 6(5), 461 - 6 Five years of experience with the exclusive use of amikacin in a neonatal intensive care unit; Powell KR et al.; From November 1, 1980, to December 31, 1985, 3959 infants were admitted to the neonatal intensive care unit and 2385 infants (60%) received 2791 courses of aminoglycoside therapy . Aminoglycoside use totaled 16,279 patient days of which 16,070 (98.7%) were with amikacin . From November 1, 1980, to January 31, 1983, 1017 pairs of pre- and posttreatment endotracheal or pharyngeal specimens yielded 318 Gram-negative bacteria isolates . From November 1, 1980, to December 31, 1985, Gram-negative bacteria were isolated from 381 clinical specimens . Of the 318 surveillance and 380 clinical isolates tested, 285 (90%) and 358 (94%), respectively, were susceptible to amikacin . Amikacin resistance did not increase during the study . Amikacin-resistant organisms were isolated more frequently from patients receiving multiple courses than those receiving single courses of amikacin and resistant organisms were not usually found before the administration of amikacin . None of the 15 amikacin-resistant isolates made 6'-N-aminoglycoside acetyltransferase and 3 isolates took up only small amounts of radiolabeled amikacin, suggesting that resistance was due to decreased permeability . The extensive use of amikacin in a neonatal intensive care unit for over 5 years did not result in an increase of amikacin-resistant Gram-negative bacteria. Am J Physiol, 1987 May, 252(5 Pt 2), R809 - 21 Ion transport in circulatory and/or septic shock; Sayeed MM; This review surveys investigations of membrane ion transport in animals in hemorrhagic, endotoxic, or bacteremic shock . The focus of the review is on ion transport studies in the skeletal muscle and liver . Skeletal muscle Na+-K+ transport alterations have been shown during the induction of shock via hemorrhage, endotoxin, or live Gram-negative bacteria in the rodent, canine, and primate species . These alterations include impairment of active cellular K+ accumulation, increased permeability to Na+ and Cl-, and membrane depolarization . The ion transport alterations in the skeletal muscle are compatible with movement of extracellular fluid into the intracellular compartment . Such fluid movements can potentially lead to decreases in circulating plasma volume and thus to circulatory deficits in shock . Studies in the liver of rats subjected to hemorrhagic or endotoxic shock indicated the failure of electrogenic Na+ pump . Although the hepatic cellular membrane permeability to Na+ relative to permeability to K+ appeared unaltered in hemorrhagic shock, endotoxic shock caused an increase in permeability to Na+ . Hepatic cellular Ca+ regulation also appeared to be adversely affected during endotoxic shock . Alterations in hepatic Na+-K+ transport and Ca+ regulation could contribute to impairment in hepatic glucose production during shock . Although mechanisms of altered membrane ion transport during shock states remain unknown, such changes could occur prior to any substantial loss of cellular metabolic energy. Int J Clin Pharmacol Ther Toxicol, 1987 May, 25(5), 282 - 8 Use of ceftazidime in the treatment of otorhinolaryngoiatric bacterial infections; Vellucci A et al.; Within the framework of an investigation into otorhinolaryngoiatric bacterial infections in Italy conducted in 1159 patients (607 with otitis media, 354 with pharyngo-tonsillitis and 198 with sinusitis), 124 ceftazidime-treated subjects (92 with otitis media, 22 with pharyngo-tonsillitis and 10 with sinusitis) were observed . The authors report the examination of a number of microbial isolates obtained in the various forms of otorhinolaryngoiatric bacterial infection . Gram-negative organisms were found as causative agents in such diseases, particularly in otitis media . Bacterial resistance to ceftazidime was assessed in all 1159 cases and proved relatively infrequent (3.6%) and markedly less than the detected resistance to other antibiotics (10.7% resistance to cefotaxime, 35% resistance to ampicillin, 43% resistance to penicillin) . Ceftazidime, used mainly in otitis media, showed very substantial clinical efficacy with positive results in as many as 97% of cases treated, which is particularly significant, if one considers that roughly 64% of the infections were caused by "difficult" gram-negative bacteria (49% by Pseudomonas). J Clin Microbiol, 1987 May, 25(5), 856 - 8 Correlation of cerebrospinal fluid endotoxinlike activity with clinical and laboratory variables in gram-negative bacterial meningitis in children; Dwelle TL et al.; Detection of endotoxinlike activity in cerebrospinal fluid by Limulus amebocyte lysate gelation has been suggested as a useful technique for the diagnosis of gram-negative bacterial meningitis . We prospectively screened 1,503 cerebrospinal fluid specimens with a Limulus amebocyte lysate microassay . The limit of sensitivity of the assay was 0.01 ng/ml . All specimens that were positive for endotoxinlike activity were subjected to confirmatory retesting, after which 38 (86%) remained positive . Comparison with available culture results revealed that 33 of 38 specimens (86%) were culture positive; 3 of the 5 culture-negative specimens were from patients on therapy for gram-negative bacterial meningitis, and 1 was from a neonate . The overall specificity of confirmed positive tests was 99.5%, with a positive predictive value of 97.3% . There was one false-negative specimen, giving an overall sensitivity of 97.3% and a negative predictive value of 99.9% . Endotoxinlike activities of greater than or equal to 150 ng/ml correlated with present illness of less than 2 days' duration (P = 0.024), elevated cerebrospinal fluid protein (P less than 0.05), and seizures (P = 0.004); levels of greater than or equal to 3,000 ng/ml correlated with neutropenia (P = 0.032), and levels of greater than or equal to 3.2 X 10(6) ng/ml correlated with death (P = 0.001) . We conclude that the Limulus amebocyte lysate microslide gelation test has prognostic value as a sensitive, specific, simple, inexpensive screening test for gram-negative bacterial meningitis. Mol Gen Mikrobiol Virusol, 1987 May, (5), 44 - 6 {Improved method of lipopolysaccharide isolation from gram-negative bacteria}; Kul'shin VA et al.; A modification of the traditional method for lipoplysaccharide isolation from the cells of grammnegative bacteria was elaborated on the basis of extraction by the hot water solution of phenol (the method of Westfahl) . To make the method simpler and to raise the yield of the product it was proposed to use the water-phenol extract without its division for plases . The nucleic acids are eliminated by precipitation from dialyzed extract at pH 3,2-3,4 achieved by addition of acetic acid . The comparative isolation of lipopolysaccharides by the classic and modified methods has confirmed the advantages of a new technique. Proc Soc Exp Biol Med, 1987 May, 185(1), 6 - 15 The inhibition of endotoxin-induced local inflammation by LDH virus or LDH virus-infected tumors is mediated by interferon; Heremans H et al.; The footpad swelling reaction induced by local injection of S . marcescens lipopolysaccharide was found to be inhibited in mice given a transplantable tumor (TA3) or cell-free ascitic fluid from tumor-bearing mice . The tumor was shown to contain LDH virus, which is known to cause inapparent persistent infections in mice . Monoclonal antibodies directed against protein VP3 of the LDH virus could partially abrogate the anti-inflammatory effect of the TA3-ascitic fluid, and, conversely, the anti-inflammatory effect could be obtained by LDH virus isolated from the tumor and reproduced by serial passage of cell-free fluids . Inhibition of the footpad reaction was seen in the acute but not in the chronic phase of LDH virus infection, suggesting that the anti-inflammatory effect might be due to endogenous interferon (IFN) which, similarly, was only detectable in the acute phase . Newcastle disease virus, another potent interferon inducer, had a similar inhibitory effect on the footpad reactivity . Moreover, the inhibitory effect of LDH virus infection could partially be abrogated by administration of a polyclonal antibody directed against murine IFN-alpha,beta . Finally, passively administered natural murine IFN-alpha,beta or recombinant murine IFN-alpha 1 (but not recombinant murine IFN-beta) was found to cause inhibition of the footpad reaction . Since Gram-negative bacteria and their lipopolysaccharides have the ability to induce a systemic interferon response, our findings suggest that this interferon may play a modulatory role in local inflammation caused by these bacteria . Our findings also open a new perspective for interferon therapy of certain inflammatory reactions to bacterial infections. J Clin Invest, 1987 May, 79(5), 1421 - 30 Human monoclonal antibodies that recognize conserved epitopes in the core-lipid A region of lipopolysaccharides; Pollack M et al.; Epstein-Barr virus (EBV)-transformed human B lymphocytes were fused with a murine-human heteromyeloma to produce stable hybrid cell lines that secreted human monoclonal antibodies (mAbs) of the IgM class that recognized conserved epitopes in the core-lipid A region of lipopolysaccharides (LPS) . Three of the mAbs reacted with epitopes on the lipid A moiety, while a fourth recognized a determinant in the core oligosaccharide . The lipid A-specific mAbs cross-reacted with heterologous rough LPS and with lipid As released by acid hydrolysis of different intact (smooth) LPS . Carbohydrate groups in the O-side chain and core oligosaccharide of isolated, smooth LPS restricted antibody access to antigenic sites on lipid A . Yet, one lipid A-reactive mAb recognized its epitope on the surfaces of a variety of intact bacteria . These findings confirm the presence of highly conserved epitopes in the core-lipid A complex and prove the existence of human B cell clones with the potential for secreting high avidity IgM antibodies that react with these widely shared determinants . Such human mAbs might provide protective activity against disease caused by diverse gram-negative bacteria. Biochemistry, 1987 Apr 21, 26(8), 2139 - 49 Interaction of polymyxin B nonapeptide with anionic phospholipids; Kubesch P et al.; The interaction of polymyxin B nonapeptide (PMBN) and polymyxin B (PMB) with the anionic phospholipids phosphatidylserine (PS), dipalmitoylphosphatidylglycerol (DPPG), dipalmitoylphosphatidic acid (DPPA), and 1:1 mixtures (w/w) of DPPA and distearoylphosphatidylcholine (DSPC) was studied by calorimetry, electron spin resonance, and fluorescence spectrometry, electron microscopy, and fusion and leakage assays . The phase transition temperatures of DPPA and DPPG were very similar when bound to PMB or PMBN, indicating that the lipids are in a similar state when bound to the cationic peptides . Both PMB and PMBN caused the interdigitation of DPPG bilayers, suggesting that the penetration of hydrophobic side chains from a peptide bound electrostatically on the surface is sufficient to induce this phenomenon . Stopped-flow experiments revealed that PMBN and PMB induced the fusion of small unilamellar PS and large unilamellar DPPA-DSPC vesicles . The aggregation of vesicles was found to be diffusion-controlled process; the subsequent fusion took place with a frequency of 10(2)-(5 X 10(2} s-1 for small vesicles and 1-100 s-1 for large vesicles . The freeze-fracture replicas of the PMB-treated vesicles displayed 12-50-nm depressions on several superimposed bilayers, indicating the formation of stable lipid-PMB domains . Since the incubation with PMBN produced similar depressions only if the specimens were fixed, PMBN-induced domain formation seems to be a reversible rapid process . The differences in the phospholipid-peptide interactions are correlated with the differences in the physiological action of the antibiotic PMB and the nonbactericidal PMBN on the cell envelope of Gram-negative bacteria. Scand J Gastroenterol, 1987 Apr, 22(3), 313 - 20 Impact of experimental endogenous gram-negative peritonitis on the pancreas of the rat as evaluated by cationic trypsin-like immunoreactivity in peritoneal fluid and serum and by electron microscopy of pancreatic tissue; Florholmen J et al.; Endogenous gram-negative peritonitis leading to septic shock was induced in rats by a defined perforation of the coecum . Cationic trypsin-like immunoreactivity (CTLI) was measured in peritoneal fluid and serum by a radioimmunoassay method . Five, 10, and 15 h after the coecal perforation, CTLI in peritoneal fluid was significantly higher than before the coecal perforation and also higher than in the corresponding control rats . Moreover, CTLI in serum was under the same conditions significantly higher 10 and 15 h after the induction of peritonitis . Gel chromatography of peritoneal fluid and serum during peritonitis showed free CTLI and CTLI bound to both alpha-1-antitrypsin and alpha-2-macroglobulin, whereas only free CTLI could be detected in serum from control rats . These findings were accompanied by local ultrastructural changes in the acinar cells as evaluated by electron microscopy . The pathophysiologic implications of the findings are discussed. Scand J Gastroenterol, 1987 Apr, 22(3), 295 - 300 Spontaneous bacterial peritonitis in cirrhosis . Incidence, diagnosis, and prognosis; Almdal TP et al.; Among 342 consecutive patients admitted to the hospital with cirrhosis of the liver 68 (17%) had ascites and had a diagnostic paracentesis performed . Fourteen episodes of peritonitis were diagnosed in 13 patients, which is an overall incidence of peritonitis of 19% . The incidence of peritonitis was 36% in patients with hepatic encephalopathy and 10% in patients without hepatic encephalopathy (P less than 0.01) . In all except one case the infecting organism was most likely of enteric origin--that is, gram-negative or anaerobic species . The infected patients had lower mean ascites pH and higher mean ascites leukocyte and polymorphonuclear cell counts than non-infected patients . However, there was a considerable overlap between the two groups, and the diagnostic sensitivity did not exceed 65% for any of these three features . The survival of infected patients without encephalopathy was 33%, which was significantly lower (P less than 0.05) than that the 89% for non-infected patients . In patients with encephalopathy the survival was identical for infected and non-infected patients. J Antimicrob Chemother, 1987 Apr, 19(4), 467 - 73 In-vivo effects of clindamycin on polymorphonuclear leucocyte phagocytosis and killing of gram-negative organisms; Bassaris HP et al.; Polymorphonuclear leucocyte (PMNL) phagocytosis and killing were evaluated in ten healthy volunteers who had received 600 mg of clindamycin intramuscularly . Serum obtained 3 h after the administration of clindamycin significantly increased PMNL phagocytosis and killing of Gram-negative aerobic organisms . Serum obtained at 12 and 24 h after the administration of the drug did not induce a significant increase in PMNL phagocytosis and killing . The administration of clindamycin had no direct effect on the PMNLs in terms of their phagocytic and bactericidal function . These results demonstrate serum-associated augmentation of PMNL function by clindamycin in vivo which may be of potential clinical benefit in the outcome of infections. J Immunol, 1987 Apr 1, 138(7), 2143 - 8 Induction of colony stimulating factor in vivo by recombinant interleukin 1 alpha and recombinant tumor necrosis factor alpha 1; Vogel SN et al.; In response to a potent inflammatory challenge, such as Gram-negative endotoxin, a number of cytokines are induced that, in turn, mediate many of the pathophysiologic alterations associated with endotoxicity . In this study, we have observed two endotoxin-associated monokines, recombinant interleukin-1 alpha (rIL 1 alpha) and recombinant tumor necrosis factor alpha (rTNF alpha), to induce colony stimulating factor (CSF) in vivo . The CSF activities produced in response to rIL 1 alpha or rTNF alpha gave rise to a mixture of granulocyte-macrophage colonies and were induced in a dose- and time-dependent fashion, peaking within 3 hr of cytokine injection (preceding peak CSF induction by endotoxin by several hours) . Combined injection of suboptimal concentrations of rIL 1 alpha and rTNF alpha were additive, and simultaneous injection of optimal concentrations of each failed to increase CSF levels over that observed with either cytokine alone . Unlike endotoxin, neither cytokine induced interferon in vivo . These findings extend our understanding of the cytokine cascade that is operative in an inflammatory response and may account for many of the observed hematopoietic alterations that accompany inflammation. Orthop Rev, 1987 Apr, 16(4), 246 - 54 A review of antibiotic prophylaxis for open fractures; Antrum RM et al.; A review of the literature strongly supports the use of antibiotic prophylaxis in open fracture management . Because of their broad spectrum of activity, cephalosporins are the drugs of choice in cases of orthopaedic trauma . The extent of the injury determines the appropriate agent and the length of time it should be given . Patients with Type 1 or Type 2 fractures require only brief treatment (one preoperative and two postoperative doses) with an agent that has good antistaphylococci activity . Cephalothin, cefazolin, cefamandole or cefuroxime are recommended . Type 3 fractures present a much greater problem because of the risk of gram-negative infection . The marked activity of cefuroxime, cefamandole, and cefotaxime against these organisms recommends their use in such cases . Combination regimens with aminoglycosides require further study to define added efficacy and to define appropriate dosing regimens . We advocate that antibiotic prophylaxis should be given as soon as possible to all patients with open fractures and Type 3 fractures, and that prophylaxis should continue until 48 hours after adequate soft tissue coverage is achieved. Acta Chir Scand, 1987 Apr, 153(4), 287 - 90 Splenectomy in the rat . Immediate and late effects on clearance of gram-negative bacteria and on monocyte activation as expressed by thromboplastin synthesis; Almdahl SM et al.; Young rats (weight 190-200 g) were subjected to splenectomy or sham laparotomy and gram-negative bacteraemia was induced by caecal perforation or by injection of viable Escherichia coli intraperitoneally (10(9) bacteria) or intravenously (2 X 10(8} . Clearance of bacteria was significantly less in the splenectomized than in the sham-laparotomized rats, irrespective of the mode of microbial inoculation . Monocyte thromboplastin activity, shown to be a sensitive indicator of gram-negative bacterial presence, was heightened in the asplenic rats . When the infectious challenge was made 15 weeks postoperatively, however, no significant difference in bacterial clearance or in monocyte thromboplastin values was found between splenectomized rats and controls . Nor was increased susceptibility to gram-negative bacteria found in asplenic rats when both operation (splenectomy vs . sham laparotomy) and bacterial challenge were performed at a late age (weight 530-580 g) . The postsplenectomy clearance of gram-negative bacteria thus seemed to be age-dependent. Acta Chir Scand, 1987 Apr, 153(4), 283 - 6 Effect of antibiotics on gram-negative sepsis in the rat . Lack of endotoxin burst; Almdahl SM et al.; Endotoxin and monocyte thromboplastin activity were evaluated in rats with gram-negative septicaemia induced by caecal perforation or intravenous Escherichia coli challenge and treated with antibiotics or placebo . Endotoxin burst was not detected in either form of septicaemia during antibiotic treatment . Thromboplastin synthesis in monocytes is known to be stimulated by endotoxin, but the rats showed no increase of monocyte thromboplastin activity after antibiotic treatment, which constituted further evidence against the concept of massive endotoxin liberation during antibiotic therapy for gram-negative septicaemia. Genetics, 1987 Apr, 115(4), 619 - 25 Mode of replicon fusion mediated by the duplicated insertion sequence IS21 in Escherichia coli; Reimmann C et al.; The insertion sequence IS21 (2.1 kb) originating from the broad-host-range IncP plasmid R68 transposes infrequently; by contrast, the IS21 tandem repeat found on the derivative R68.45 is highly active in transpositional mobilization of other replicons in a variety of Gram-negative bacteria . The mobilized plasmids are joined to R68.45 by single IS21 copies in direct orientation . The formation of IS21 tandem duplications was observed in cointegrates between R68.45 and pBR325::IS21 and also in an RP1::IS21 plasmid derivative in which a segment located between two directly repeated copies of IS21 was deleted spontaneously . We speculate that IS21 tandem repeats can arise when the termini of two IS21 elements are specifically joined in a transposition or deletion event . A resistance gene flanked by two IS21 elements in direct orientation did not behave as a transposon . The omega fragment carrying transcription and translation stop signals was inserted into various sites of the IS21 tandem repeat; in this way it could be shown that the left IS21 element (which is next to the kanamycin resistance gene in R68.45) was 100 times more active in cointegrate formation than was the right-hand element . Cointegrates between the conjugative plasmid R751 and pBR325 derivatives carrying IS21 and IS21::omega in tandem contained a single IS21 at one replicon junction and a single IS21::omega at the other . In the IS21 duplications the inner IS21 ends were preferentially recognized (presumably by IS21 transposase), whereas the outer termini were not required for cointegrate formation . Based on these findings a conservative (simple) pathway of transposition is proposed for R68.45 and other plasmids with an IS21 tandem repeat.(ABSTRACT TRUNCATED AT 250 WORDS) Infect Immun, 1987 Apr, 55(4), 916 - 22 Purification and characterization of serotype 6 fimbriae from Bordetella pertussis and comparison of their properties with serotype 2 fimbriae; Cowell JL et al.; Fimbriae were removed from Bordetella pertussis (serotype 1.3.6) by mechanical shearing and purified by precipitation with ammonium sulfate, pH-dependent precipitation at pH 7.4, followed by two successive extractions of the precipitated fimbriae with 4 M urea . By electron microscopy, the precipitated fimbriae appeared as aggregated bundles of long, relatively straight filaments which were disaggregated to individual flexuous filaments at pH 10.5 . These purified fimbriae were identified as serotype 6 agglutinogens, since antibody to the purified fimbriae agglutinated B . pertussis strains serotyped as 1.3.6, 1.2.3.6, or 1.2.3.4.6 but did not agglutinate strains of serotype 1.2.3.4, 1.2.3, or 1.3 . In contrast, antibody to serotype 2 fimbriae only agglutinated B . pertussis strains containing serotype 2 agglutinogen . Purified type 6 and 2 fimbriae were found to be weakly cross-reactive by enzyme-linked immunosorbent assay, using polyclonal antibody to each type of fimbria . In an immunoblot assay, polyclonal antibodies to a 22,000-dalton subunit of fimbriae from B . bronchiseptica reacted strongly with the type 2 fimbrial subunit of B . pertussis, but only weakly with the type 6 subunit . When subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the protein subunit of the type 6 fimbriae migrated with a molecular weight of 21,500, whereas the type 2 fimbrial subunit had a molecular weight of 22,000 . The two types of subunits had similar amino acid compositions and showed amino-terminal sequence homology in 15 of 21 amino acids . The amino-terminal amino acid sequences of the B . pertussis fimbriae were distinct from those reported for fimbriae from other gram-negative bacteria . Neither the type 6 nor the type 2 fimbriae caused hemagglutination when assayed with several types of erythrocytes. Biochem Biophys Res Commun, 1987 Mar 30, 143(3), 1063 - 8 A 2-deoxy analogue of KDO as the first inhibitor of the enzyme CMP-KDO synthetase; Claesson A et al.; The two KDO analogues 2,6-anhydro-3-deoxy-D-glycero-D-galacto-octonate and 2,6-anhydro-3-deoxy-D-glycero-D-talo-octonate were synthesized and tested as inhibitors of the enzyme CTP:CMP-deoxyoctulosonate cytidylyltransferase (CMP-KDO synthetase) from Gram-negative bacteria . Only compound 4, the 2-deoxy analogue of beta-KDO-pyranose, was found to be an inhibitor with a Ki of 3.9 microM. Biochim Biophys Acta, 1987 Mar 19, 923(3), 463 - 9 Purification and properties of a pepstatin-insensitive carboxyl proteinase from a gram-negative bacterium; Oda K et al.; A carboxyl proteinase was found in the culture filtrate of a Gram-negative bacterium . The optimum for the action of the purified enzyme was approx . pH 3 and its caseinolytic activity was not inhibited by carboxyl proteinase inhibitors, such as pepstatin, Streptomyces pepsin inhibitor and diazoacetyl-DL-norleucine methyl ester . 1,2-epoxy-3-(p-nitrophenoxy)propane modified the enzyme with concomitant loss of its enzyme activity . The enzymatic and physicochemical properties of the enzyme were compared with those of known pepstatin- and diazoacetyl-DL-norleucine methyl ester-insensitive carboxyl proteinases previously reported . To our knowledge, this is the first carboxyl proteinase isolated from bacteria. Am Rev Respir Dis, 1987 Mar, 135(3), 671 - 5 Increased salivary elastase precedes gram-negative bacillary colonization in postoperative patients; Dal Nogare AR et al.; The upper airway epithelium is coated with fibronectin, a glycoprotein that covers receptor sites for gram-negative bacteria and prevents them from colonizing the oropharynx . We investigated the identity of salivary proteolytic enzymes capable of degrading fibronectin in a group of 16 patients who had elective cardiac surgery . Six patients became colonized with gram-negative bacteria (Group C) and 10 did not (Group NC) . Salivary elastase activity was low in both groups preoperatively . Twenty-four hours after surgery, salivary elastase activity increased in Group C, and it remained elevated at 48 and at 72 h . Fibronectin digestive activity of the saliva of patients in Group C was also increased within 24 h of surgery, and salivary elastase and fibronectin digestive activity were highly correlated (r = 0.86, p less than 0.001) . Enzyme inhibition experiments showed that most of the fibronectin digestive activity was due to elastase from polymorphonuclear cells (PMN), and the molecular weight of the salivary enzyme digesting fibronectin was 30,000 daltons (similar to the molecular weight of elastase) . Levels of antileukoprotease, the major elastase inhibitor in saliva, were normal in patients with increased elastase activity . We conclude that salivary elastase is of PMN origin, increases prior to gram-negative bacillary colonization of the pharynx, and is responsible for most of the fibronectin digestive activity of the saliva. EMBO J, 1987 Mar, 6(3), 823 - 31 Signal peptide amino acid sequences in Escherichia coli contain information related to final protein localization . A multivariate data analysis; Sjostrom M et al.; With few exceptions, the signal peptides from proteins inserted into, or translocated through, the membranes of gram-negative bacteria or the endoplasmic reticulum of eukaryotes have no sequence homologies . Therefore these signal peptides have not been considered to contain information related to the different final localizations of the proteins . In this study, 43 signal peptide amino acid sequences from proteins with different final localizations in Escherichia coli have been subjected to a multivariate data analysis . Each amino acid residue was characterized by 20 physico-chemical properties, yielding a multivariate property profile for each peptide . The similarities/dissimilarities in the property profiles for the signal peptides from different classes were compared with each other by generating few-dimensional partial least squares (PLS) discriminant plots . With this approach, signal peptides from proteins localized to the periplasmic space (PS), the outer membrane (OM), and the extracellular surroundings (excreted proteins), were separated into distinct groups . Signal peptides from pili proteins were not separated from the OM signal peptides and only partly from the PS signal peptides, but were clearly different from the signal peptides of the excreted proteins . Signal peptides from inner membrane proteins were similar to those of the PS peptides . The size and the hydrophobicity of different peptide segments were responsible for the separation of the signal peptide classes . For example, the hydrophobicity of the N-terminal segment of the signal peptides increased with an increased distance from the cytoplasm of the final localization for the corresponding proteins . Thus, many signal peptides from proteins with different final localizations in E . coli have different discernible physico-chemical profiles. Infect Immun, 1987 Mar, 55(3), 668 - 73 Prophylactic administration of interleukin-2 protects mice from lethal challenge with gram-negative bacteria; Chong KT; Prophylactic administration of recombinant human interleukin-2 (IL-2) in mice enhanced survival and produced complete recovery from an otherwise lethal acute bacterial infection . IL-2 was administered as a single intraperitoneal or intravenous bolus dose to CDI mice 18 h before challenge with a lethal dose of a clinical isolate of Escherichia coli type O2 (minimal 100% lethal dose, 6 X 10(7) CFU per mouse) . At IL-2 dosages of 7 X 10(6) U/kg, 90% of treated CDI mice survived as compared to 0% for the excipient buffer control animals (P less than 0.001) . This protective effect was also demonstrable in immune-deficient beige mice . The IL-2 effect was dose dependent; protection was consistently observed in mice pretreated with IL-2 at doses ranging from 1.8 X 10(6) to 7 X 10(6) U/kg . However, at 3.5 X 10(5) U/kg the protective effect was more variable . The route of administration of IL-2 was shown to play an important role; when IL-2 and challenge bacteria were given by the same route (either intravenously or intraperitoneally), protection was readily observable, but when IL-2 and challenge bacteria were given by different routes, little or no protective effect was observed . The protective effect was fully inducible as early as 1 h after IL-2 administration and was effective against various strains of gram-negative bacteria, indicating that the probable mode of action represents control of the establishment of infection by increased activity of the nonspecific host defense mechanisms . The IL-2 effect was abrogated by the administration of carrageenan, suggesting a possible role of macrophages . These data demonstrate that IL-2 may be a potentially useful adjunct for the prophylaxis of bacterial infections in both clinical and veterinary medicine. Gan To Kagaku Ryoho, 1987 Mar, 14(3 Pt 1), 674 - 9 {Cefmenoxime or piperacillin plus amikacin . A prospective randomized comparison of empiric antibiotic therapy of febrile granulocytopenic cancer patients}; Sampi K et al.; Cefmenoxime plus amikacin was compared in a prospective randomized trial with our standard regimen of piperacillin plus amikacin as an empiric therapy for fever in patients with granulocytopenia . Initial profound granulocytopenia (fewer than 100/mm3 mature granulocytes) was present in approximately 45% of the patients in trial of both treatment groups . Of 53 microbiologically and clinically documented infections treated with piperacillin plus amikacin, 36 (68%) showed improvement . Of 48 microbiologically and clinically documented infections treated with cefmenoxime plus amikacin, 23 (48%) showed improvement . The response rate for gram-negative infections treated with cefmenoxime plus amikacin was lower than that for infections treated with piperacillin plus amikacin . Toxicity was minimal, with an equivalent incidence of skin rash, diarrhea and hepatic dysfunction . Although clinical efficacy of the combination of piperacillin plus amikacin may be superior to cefmenoxime plus amikacin therapy, this study demonstrated no statistically significant differences. Infection, 1987 Mar-Apr, 15(2), 146 - 52 {Antibodies to lipoid A in the treatment of septic shock}; Jaspers L et al.; The protective effect of high-titer anti-lipid A hyperimmune globulin with respect to the course of the disease and the mortality rate was studied in patients with septicemia verified by positive blood cultures . Six patients were treated with anti-lipid A in an open study . Dramatic improvement in fever curves and clinical condition in some of the patients encouraged us to start a randomized double blind study . So far, 17 patients have entered the study, 16 of whom were evaluable . Immediately after a positive blood culture was found, patients received either high doses of anti-lipid A or placebo (saline solution) on two subsequent days . Before and after each infusion blood samples were taken in order to assess serum bactericidal activity and anti-lipid A titers . Because of the still small numbers of patients the results of both studies were summarized . In all patients treated with anti-lipid A clear-cut increases in anti-lipid A titers were shown . Patients with repeated gram-negative infections showed higher median anti-lipid A titers than patients without such a history . The patients treated with anti-lipid A immune globulin ran a significantly milder course than the placebo group . The severe signs of septic shock were reversed in seven of 15 patients on anti-lipid A compared to two of seven patients treated with placebo . In the anti-lipid A-treated group, three of 15 patients died, and in the placebo group two of seven . This difference is not statistically significant. Zentralbl Bakteriol Mikrobiol Hyg {B}, 1987 Mar, 183(4), 337 - 57 {Bacteriological, clinical, and pharmacokinetic studies of perioperative antibiotic prophylaxis in head and neck surgery}; Borneff M et al.; The point of this study was to analyze the possible benefits of perioperative antibiotic prophylaxis in patients from whom oral cavity or throat tumors are removed . The criteria used to judge the efficacy of each treatment included the clinical course of the treatment, the bacterial colonization of the surgical area as well as the growth of bacteria during the postoperative phase . 50 patients were chosen and grouped according to their surgical treatment: laryngectomy (n = 20), partial laryngectomy (n = 22) or tongue, floor of the mouth, soft palate, gum or base of the tongue partial resection (n = 8) . Within each surgical group, patients were randomly chosen for antibiotic prophylaxis; others constituted the untreated control group . The antibiotic prophylaxis consisted of 5 g Mezlocillin administered at the time of narcosis for 20 min followed by 0.5 g Metronidazol for 10 min . These medications were given in 8-hour intervals for three days following surgery . Investigation of the first 20 patients (prophylaxis group n = 7, control group n = 13) revealed that the combination of Mezlocillin and Metronidazol positively influenced post-operative recovery (no complications) while the patients without prophylactic antibiotic treatment suffered general or local complication leading to, in 10 cases, the necessity of postoperative therapy . On the basis of these results, the random grouping of the patients was ended and all 30 remaining patients were given the antibiotic prophylaxis . Regardless of antibiotic treatment, the great majority of microbes isolated from throat swabs and tracheal secretions were gram-negative, aerobic bacteria . A prerequisite for efficacious prophylaxis is that the antibiotics be applied before the operation, so that a sufficient concentration is present at the time of pharyngotomy . On the basis of pharmacokinetic investigations, administration of the antibiotic 30 min preoperatively fulfills this requirement . Further, our recommendation, based on our measurement of the spectrum of bacteria present and their growth is that the antibiotics be applied over a period of three days postoperatively . This recommendation is also based on the fact that some patients (those having undergone partial laryngectomy or tongue, floor of the mouth, base of the tongue partial resections) have suffered loss of the swallowing reflex so that there exists a continuous contamination of the surgical area with pathogens or facultative pathogens coming from the nasal or oral cavities. J Clin Invest, 1987 Mar, 79(3), 918 - 25 Protein C prevents the coagulopathic and lethal effects of Escherichia coli infusion in the baboon; Taylor FB Jr et al.; Gram-negative septicemia elicits multiple abnormalities of the coagulation system . Although products of coagulation can lead to clot formation, thereby potentiating organ damage, recent work has shown that low concentrations of thrombin can protect animals from the shock state . Because these amounts of thrombin also lead to formation in vivo of the anticoagulant enzyme, activated protein C, we examined the role of protein C in modulation of Escherichia coli shock in baboons . First, we infused activated protein C and lethal concentrations of E . coli organisms, which prevented the coagulopathic, hepatotoxic, and lethal effects of E . coli . Second, using an antibody to protein C we blocked protein C activation in vivo to determine if this influenced the response to lethal and sublethal concentrations of E . coli organisms . Under these conditions the response to lethal concentrations of E . coli organisms was made more severe and the response to sublethal concentrations of E . coli was made lethal . The coagulopathic, hepatotoxic, and lethal responses in this latter case were prevented by infusion of exogenous protein C. J Virol, 1987 Mar, 61(3), 812 - 8 Macrophages from endotoxin-hyporesponsive (Lpsd) C3H/HeJ mice are permissive for vesicular stomatitis virus because of reduced levels of endogenous interferon: possible mechanism for natural resistance to virus infection; Vogel SN et al.; The C3H/HeJ mouse strain bears an autosomal gene defect, Lpsd, which results in a greatly diminished capacity to respond to endotoxin, the ubiquitous lipopolysaccharide derived from the cell walls of gram-negative bacteria . These mice also exhibit greater susceptibility to a variety of viral and bacterial infections than syngeneic, fully lipopolysaccharide-responsive (Lpsn) mouse strains and possess macrophages with defects in differentiation which are reversed by treatment with exogenous interferon (IFN) . To test directly the hypothesis that C3H/HeJ macrophages are deficient in endogenous IFN levels, macrophages from C3H/HeJ (Lpsd) and C3H/OuJ (Lpsn) mice were compared for sensitivity to vesicular stomatitis virus . At a multiplicity of infection of 0.1, C3H/OuJ macrophages were completely refractory to infection, whereas C3H/HeJ macrophages were permissive for replication, and infection resulted in 100% cytopathic effect . These findings were confirmed with a second inbred Lpsn and Lpsd strain pair . Levels of 2',5'-oligoadenylate synthetase were significantly higher in Lpsn cells . C3H/HeJ macrophages, derived from bone marrow precursors under the influence of macrophage colony-stimulating factor, shown previously to induce IFN in macrophages, were as refractory as C3H/OuJ macrophages . Exposure of nonpermissive macrophages to anti-IFN-alpha/beta antibody prior to infection rendered cells permissive . Our findings suggest that endotoxin provides a primary stimulus for the maintenance of normal macrophage differentiation and innate resistance via the induction of endogenous IFN by macrophages. Eur J Biochem, 1987 Feb 16, 163(1), 73 - 7 Nucleotide sequence of the gene for the peptidoglycan-associated lipoprotein of Escherichia coli K12; Chen R et al.; During attempts to clone the gene for the Escherichia coli outer membrane protein III another gene was recovered . Its nucleotide sequence was determined and the deduced amino acid sequence showed that the gene does not encode protein III . It codes for a 173-residue polypeptide; 21 NH2-terminal residues are typical for a signal peptide . The sequence around the putative site (Ala-Cys) for removing this peptide, Ala-Ile-Ala-Ala-Cys-Ser-Ser-Asn, is highly homologous to that of the major cell envelope lipoprotein (Braun lipoprotein) surrounding its processing site; it is also homologous to the consensus pentapeptide Leu-Leu-Ala-Gly-Cys present in other lipoproteins of gram-negative bacteria . It could be shown that the gene expresses a lipoprotein with all the properties, including the amino acid composition, of the peptidoglycan-associated lipoprotein (PAL) {Mizuno, T . (1979) J . Biochem . (Tokyo) 86, 991-1000} . Therefore, the cloned gene is the pal gene . The protein does not contain hydrophobic regions which would serve as a membrane anchor . Tandemly repeated amino acid sequences exist at and near the NH2-terminus of the mature protein which are homologous to such repeats in the Braun lipoprotein, suggesting a common origin of this part of the two proteins . Attempts to place a transposon into the pal gene were unsuccessful . Hence the complete absence of the protein may be lethal and its function remains unknown. Arch Surg, 1987 Feb, 122(2), 207 - 12 Effect of altered volume of distribution on aminoglycoside levels in patients in surgical intensive care; Niemiec PW Jr et al.; The apparent volume of distribution (Vd) of aminoglycosides was found to be increased in 100 patients in a surgical intensive care unit who had gram-negative pneumonia or intraabdominal sepsis and acute physiologic scores greater than 12 . Following loading or maintenance doses, carefully timed blood samples were collected for measurements of serum concentrations by fluorescence polarization immunoassay . The Vd, determined by linear regression analysis of a one-compartment model using the Sawchuk-Zaske method, was 0.34 +/- 0.121 L/kg and was larger than the normal Vd of 0.20 to 0.25 L/kg, suggesting a 36% to 70% increase in extracellular fluid volume . Since there is a predictable increase in aminoglycoside Vd in the septic surgical patient, a proportionately larger aminoglycoside dosage is required initially to achieve desirable peak serum levels . Close monitoring of blood levels during maintenance dosing is suggested since dynamic changes in renal function and aminoglycoside Vd occur in the critically ill. Blood, 1987 Feb, 69(2), 652 - 9 Cellular and subcellular localization of the bactericidal/permeability-increasing protein of neutrophils; Weiss J et al.; Human and rabbit polymorphonuclear leukocytes contain a bactericidal/permeability-increasing protein (BPI), a potent cytotoxin active specifically against gram-negative bacteria . To identify the cell population(s) producing BPI, we have examined mature and immature human blood cells for BPI by immunofluorescence of intact cells and radioimmunoassay and bioassay of cell extracts . By immunofluorescence and radioimmunoassay of cells from peripheral blood, BPI was detected only in neutrophils; immunofluorescent staining was punctate, indicative of the granule localization of BPI . Nearly all (greater than 90%) BPI was recovered during the subcellular fractionation of neutrophils (N2 cavitation and discontinuous Percoll gradient) in fractions containing primary granules . Little BPI was released from intact cells during degranulation (cytochalasin B and f-Met-Leu-Phe) or could be extracted from isolated granules with salt or weak acid, which suggests that most granule-associated BPI is membrane bound . Double staining of bone marrow smears for BPI and lactoferrin revealed BPI only in neutrophil precursors including (pro)myelocytelike cells lacking lactoferrin, a marker of neutrophil secondary granules . Of several human cell lines tested, only the promyelocytelike HL-60 (and to a lesser extent, KG-1) cells contained BPI . BPI was present in a more mature subpopulation (less than 25%) of untreated HL-60 cells, recognized by surface marker analysis (rosetting with IgG-sensitized sheep RBC, the absence of proliferation-associated cell surface antigen) . Induction of neutrophilic or monocytic differentiation caused, respectively, a small (approximately 50%) rise or fall in the BPI content . These findings indicate that BPI is a specific product of the neutrophil lineage and, hence, of the specialized cytotoxic apparatus of the neutrophil that plays an essential role in host defense v gram-negative bacteria. Blood, 1987 Feb, 69(2), 640 - 4 Recombinant tumor necrosis factor causes activation of human granulocytes; Larrick JW et al.; We have tested the hypothesis that tumor necrosis factor (TNF), by binding to and activating granulocytes, may contribute to the pathogenesis of gram-negative sepsis and the adult respiratory distress syndrome (ARDS) . Buffy coat granulocytes incubated with as little as 0.5 ng/mL of recombinant TNF (rTNF) showed a dose-related increase in nitroblue tetrazolium dye reduction, in granulocyte polarization, in superoxide anion release, and in visually apparent aggregation . Purified lipopolysaccharide (1 microgram/mL) caused polymorphonuclear (PMN) aggregation and activation that was neutralized by polymyxin B . The release of superoxide was augmented by preincubation of the PMNs with gamma-interferon . The effect of TNF was neutralized by TNF-specific murine monoclonal antibodies but not by polymyxin B . Scatchard analysis of 125I-rTNF binding to granulocytes revealed about 1,200 receptors per cell with a Kd of 4.9 X 10(-10) mol/L . These results suggest that the release of TNF by mononuclear phagocytes contributes to granulocyte activation and aggregation during inflammation. Antimicrob Agents Chemother, 1987 Feb, 31(2), 300 - 5 Molecular hybridization versus isoelectric focusing to determine TEM-type beta-lactamases in gram-negative bacteria; Jouvenot M et al.; Isoelectric focusing and molecular hybridization with a TEM DNA probe were used to screen for TEM beta-lactamase in 328 bacterial isolates representing 11 gram-negative genera . The TEM enzyme was detected in 50% of isolates, and nine additional types of beta-lactamase could be identified in 36.9% of isolates . The TEM gene was detected in 53.6% of isolates . The results obtained by both methods were concordant in 92.7% of the entire sample . In situ colony hybridization with a specific probe therefore appears to be a convenient method to screen rapidly for the presence of homologous genetic sequences among a large number of isolates . Positive hybridization was observed for 16 isolates in which no TEM beta-lactamase was detected by isoelectric focusing . The significance of this hybridization remains to be determined. Environ Res, 1987 Feb, 42(1), 63 - 71 Toxicologic interactions between ozone and bacterial endotoxin; Peavy DL et al.; The effects of acute exposure of mice to bacterial lipopolysaccharide (LPS), the endotoxin of gram negative microorganisms, and ozone (O3) have been investigated . Intraperitoneal (ip) administration of 5 mg/kg LPS to CD-1 mice followed by exposure to 15 ppm O3 for 1.5 hr produced synergistic effects as measured by pulmonary edemagenesis and lethality assays . In contrast, ip administration of 0.1-1.6 mg/kg LPS to CD-1 mice over 5 consecutive days, a dose regimen resulting in LPS tolerance, protected against a lethal challenge of 20 ppm O3 for 3 hr . A statistically significant increase in catalase and glutathione peroxidase activity was measured in homogenates of lungs obtained from CD-1 mice receiving a tolerance-inducing regimen of LPS . These results demonstrate that two, distinct toxicologic interactions can occur between O3 and bacterial LPS . Synergism between these agents could explain, in part, the increased susceptibility of O3-exposed animals to respiratory infection with gram negative microorganisms . Protection resulting from LPS-induced increases in pulmonary antioxidant activity provides additional evidence that O3 and, possibly, LPS mediate their toxicity through oxidative mechanisms. Bone Marrow Transplant, 1987 Feb, 1(3), 271 - 9 Autologous bone marrow transplantation for patients with acute myeloid leukaemia and acute lymphoblastic leukaemia--a comparison; Anderson CC et al.; Forty-two patients with acute leukaemia were treated with autologous bone marrow transplantation (ABMT) using a combination chemotherapy protocol for bone marrow ablation . The response to high-dose chemotherapy and ABMT and its associated morbidity and mortality have been compared in 24 patients with acute myeloid leukaemia (AML) and 18 patients with acute lymphoblastic leukaemia (ALL) . In 16 patients with AML treated with ABMT during first complete remission (CR), ten patients (62.5%) remain in unmaintained remission; median follow up is 32 months . In eight patients with ALL treated in first CR, only one remains in remission 32 months post-ABMT, with three patients dying non-leukaemic deaths . Fourteen of 18 patients (AML and ALL) treated after first remission have died of recurrent leukaemia, two died non-leukaemic deaths and two remain well 31 and 55 months post-ABMT; both have ALL . The length of hospital stay and the amount of blood product support were similar in both groups . Haematological recovery post-ABMT was delayed in patients with AML compared to patients with ALL but this difference was not significant . Rapidly progressive lung infection was thought to be the cause of four early deaths (4/18) in patients with ALL but none in patients with AML . Severe gram-negative infections were significantly more common in patients with AML. Immunol Lett, 1987 Feb, 14(3), 203 - 8 Antibody-independent killing of gram-negative bacteria via the classical pathway of complement; Loos M et al.; The experiments in this paper provided evidence that, besides lipopolysaccharides (LPS), porins of gram-negative bacteria bind to C1q and C1 . From these experiments, we concluded that the association of LPS and porins (outer membrane proteins, OMP) may potentiate the C1q and C1 binding in the absence of specific antibodies . This antibody independent binding of C1 to LPS and porins is a prerequisite for the activation of the classical pathway of complement leading to the killing of serum-sensitive bacteria. Antimicrob Agents Chemother, 1987 Feb, 31(2), 306 - 11 Organization of two sulfonamide resistance genes on plasmids of gram-negative bacteria; Swedberg G; The organization of two widely distributed sulfonamide resistance genes has been studied . The type I gene was linked to other resistance genes, like streptomycin resistance in R100 and trimethoprim resistance in R388 and other recently isolated plasmids from Sri Lanka . In R388, the sulfonamide resistance gene was transcribed from a promoter of its own, but in all other studied plasmids the linked genes were transcribed from a common promoter . This was especially established with a clone derived from plasmid R6-5, in which transposon mutagenesis showed that expression of sulfonamide resistance was completely dependent on the linked streptomycin resistance gene . The type II sulfonamide resistance gene was independently transcribed and found on two kinds of small resistance plasmids and also on large plasmids isolated from clinical material. J Biol Chem, 1987 Jan 25, 262(3), 1122 - 8 The biosynthesis of gram-negative endotoxin . A novel kinase in Escherichia coli membranes that incorporates the 4'-phosphate of lipid A; Ray BL et al.; Extracts of Escherichia coli contain an enzyme that generates the beta,1----6 linkage of lipid A from fatty-acylated monosaccharide precursors, according to the reaction: 2,3-diacyl-GlcN-1-P + UDP-2,3-diacyl-GlcN----2,3-diacyl-GlcN (beta, 1----6)2,3-diacyl-GlcN-1-P + UDP (Ray, B . L., Painter, G., and Raetz, C . R . H . (1984) J . Biol . Chem . 259, 4852-4859) . We now describe a membrane-bound kinase that phosphorylates the 4'-position of the above tetraacyldisaccharide 1-phosphate product . The lipid A 4'-kinase is distinct from the diglyceride kinase of E . coli . When crude membrane preparations are employed, several nucleoside triphosphates are able to support the phosphorylation of the tetraacyldisaccharide 1-phosphate, but ATP is the most efficient . The 4'-kinase requires Mg2+ and is stimulated by phospholipids, especially cardiolipin . Under optimal conditions the specific activity in crude extracts is 0.5 nmol/min/mg . The enzyme is rapidly inactivated by preincubation in the presence of detergents, such as Nonidet P-40 or octylglucoside, but phosphoenolpyruvate and glycerol stabilize the enzyme . The product generated in vitro has been characterized by fast atom bombardment mass spectrometry and by 1H and 31P NMR spectroscopy . Those analyses confirm that the 4' hydroxyl is the site of phosphorylation . The 4'-kinase reported here is likely to represent a key step in the de novo biosynthesis of lipid A. Br Med J (Clin Res Ed), 1987 Jan 24, 294(6566), 208 - 10 Non-secretion of ABO blood group antigens as a host susceptibility factor in the spondyloarthropathies; Shinebaum R et al.; Gram negative bacteria precipitate reactive arthritis and may be concerned in the pathogenesis of ankylosing spondylitis and other spondyloarthropathies . Susceptibility to many infectious agents is associated with ABO blood group or secretor state, or both . The distribution of the ABO blood group or secretor state, or both, was therefore determined in 87 patients with ankylosing spondylitis and 32 with other forms of spondyloarthropathy . The prevalence of non-secretors was significantly increased in the total patient group (54/114; 47%) and in the subgroup with ankylosing spondylitis (41/84; 49%) compared with local controls (89/334; 27%) (p less than 0.001) . Other subgroups of patients showed a similarly increased prevalence of non-secretion (33-47%) . The distribution of ABO blood groups did not differ between patients and controls . The association between non-secretor state and ankylosing spondylitis strengthens the hypothesis that ankylosing spondylitis is a form of reactive arthritis . It also suggests several pathogenic mechanisms which may be relevant to the initial hostparasite interaction in ankylosing spondylitis. Schweiz Med Wochenschr, 1987 Jan 17, 117(3), 84 - 90 {Indications for passive immunotherapy in infectious diseases}; Baumgartner JD; The most important indications for passive immunotherapy in the field of infectious diseases are reviewed . Intramuscular immunoglobulins are useful in prophylaxis and treatment of diseases due to bacterial exotoxins and in prophylaxis of some viral infections . However, their efficacy against bacterial infections has not been demonstrated . Intravenous immunoglobulins have theoretical advantages in these infections . The few clinical studies performed in neonates and in patients of surgical intensive care units have suggested modest benefits . However, these studies did not allow to decide whether intravenous immunoglobulins have a role in these situations . Another approach still under investigation is to administer polyclonal antibodies directed against the central part of the endotoxin, the structure of which is well preserved among gram-negative bacteria . These antibodies have improved the survival of patients with gram-negative bacteremia or septic shock . When given prophylactically, they have reduced the incidence of gram-negative shock and related mortality in patients from surgical intensive care units . Further studies are in progress to determine the class and the precise specificity of these protective antibodies. Cell, 1987 Jan 16, 48(1), 55 - 62 Biosynthesis and structure of stable branched RNA covalently linked to the 5' end of multicopy single-stranded DNA of Stigmatella aurantiaca; Furuichi T et al.; Stigmatella aurantiaca, a gram-negative bacterium, contains approximately 500 copies per cell of a short single-stranded linear DNA (multicopy single-stranded DNA: msDNA) . This DNA is attached to a branched RNA (msdRNA) by its 5' end . The entire sequence of msdRNA was determined and found to consist of 76 bases . The msDNA is linked at the 19th G residue of msdRNA by a 2', 5' phosphodiester linkage . The coding region for msdRNA (msr) is located downstream of the coding region for msDNA (msd) . These coding regions exist in opposite orientation with respect to each other and overlap by 8 bases at their 3' ends . Biosynthesis of RNA-linked msDNA was characterized and mechanisms of synthesis are proposed. Cell, 1987 Jan 16, 48(1), 47 - 53 Branched RNA covalently linked to the 5' end of a single-stranded DNA in Stigmatella aurantiaca: structure of msDNA; Furuichi T et al.; Stigmatella aurantiaca is a gliding, gram-negative bacterium that shows a spectacular fruiting body formation upon starvation of nutrient . This bacterium was found to contain approximately 500 copies per cell of a short single-stranded linear DNA (multicopy single-stranded DNA: msDNA) . The primary structure of msDNA was determined and found to consist of 162 or 163 deoxyribonucleotides . Its unique chromosomal gene was cloned and sequenced . The msDNA was found to be attached to a branched RNA by its 5' end . Structural analysis of the branched RNA revealed that it consists of a triribonucleotide, 5'A-G-(C or U)3', and that msDNA is branched out from the 2' position of the rG residue forming a 2', 5' phosphodiester linkage with the dC residue at the 5' end of msDNA. Klin Wochenschr, 1987 Jan 15, 65(2), 61 - 8 The clinical significance of prostaglandins and thromboxane as mediators of septic shock; Oettinger W et al.; An evaluation was made of 106 surgical patients with Gram-negative septic shock, both for clinical criteria as well as the biochemical mediators endotoxin, prostaglandin F2 alpha, prostaglandin I2 (prostacyclin), and thromboxane . These data were correlated to various defined shock phases, functional data of vital organs, and clinical outcome . Patients underwent invasive organ function monitoring and the usual laboratory tests of intensive care . Prostaglandins and thromboxane were measured radioimmunologically, endotoxin by the limulus amebocyte lysate test . Endotoxin proved to be a more accurate predictor of severe sepsis than did positive blood cultures . Endotoxin as well as prostaglandins and thromboxane are predominantly released in early shock phases, appearing in plasma concentrations, which correlate with the severity of organ failure . Sepsis-induced respiratory failure coincides with a deterioration of pulmonary prostaglandin inactivation, which contributes to the release mechanism . High systemic prostacyclin activity benefits the patients' organ functions and clinical outcomes, while a predominance of thromboxane seems to effect the opposite . Transpulmonary-thromboxane gradients correlate significantly with pulmonary hypertension in the early phases of septic shock. Nature, 1987 Jan 8-14, 325(7000), 179 - 80 Binding of a non-beta-lactam antibiotic to penicillin-binding proteins; Nozaki Y et al.; In the search for new beta-lactam antibiotics of natural origin, the discoveries of cephamycins and sulfazecins (monobactams) were important turning points in that they accelerated many screening efforts aimed at other new compounds . In our target-directed screening for beta-lactam antibiotics using beta-lactam hypersensitive mutants, we have examined Gram-negative bacteria isolated from natural habitats and have recently reported several types of beta-lactam antibiotics such as cephabacins and formadicins . Here we report a novel antibiotic, lactivicin, found using this system . Although lactivicin has various biological activities commonly observed in beta-lactam antibiotics, it does not possess a beta-lactam ring in its molecule, but has the unique structure of a dicyclic dipeptide. Am J Vet Res, 1987 Jan, 48(1), 91 - 5 Experimental infection and abortion of pregnant guinea pigs with a unique spirillum-like bacterium isolated from aborted ovine fetuses; Bryner JH et al.; Study was made of the pathogenicity of a spirillum-like, anaerobic, gram-negative bacterium, originally isolated from aborted lambs, for pregnant guinea pigs . Reproducible conditions for propagation and preservation of the bacterium were determined as requisite for the preparation of cultures for animal inoculation . A preliminary experiment was done with 10 pregnant guinea pigs to test for an infective dose of organisms that would produce abortion . High-passage cultures (n = 50) were used to inoculate these guinea pigs intraperitoneally . Six of 10 guinea pigs aborted, and the organism was cultured from fetal tissues of 5 guinea pigs . Isolates from 3 of the 6 guinea pigs were propagated through 4 passages on blood agar and used to infect 3 groups, each of 5 guinea pigs . A 4th group of 5 guinea pigs was inoculated with the original culture . Three of 5 animals in the first 3 groups, which had been given the low-passage cultures from the preliminary trial, and 2 of 5 guinea pigs in the 4th group, which had been given the original culture, aborted . Antibody against the spirillum was detected in 19 of 30 inoculated guinea pigs . The major microscopic lesions were acute suppurative placentitis and splenitis . This bacterium retained pathogenic properties sufficient to cause infection, abortion, and microscopic lesions in two-thirds of the guinea pigs, in spite of high in vitro passage . The organism has unique ultrastructures, and its genus and species are yet to be determined. J Infect Dis, 1987 Jan, 155(1), 113 - 8 Septicemic plague in New Mexico; Hull HF et al.; Eighteen of the 71 cases of plague reported in New Mexico from 1980 to 1984 were septicemic . We reviewed these cases to better describe the clinical presentation of this disorder and to identify risk factors for developing septicemic plague . The symptoms (fever, chills, malaise, headache, and gastrointestinal symptoms) and signs (tachycardia, tachypnea, and hypotension) of septicemic plague are similar to those of other forms of gram-negative septicemia . Abdominal pain was reported in nearly half of the cases, and differential white blood cell counts revealed a marked shift to the left . The risk of developing septicemic plague was higher for persons greater than 40 years of age . Because of empirical antibiotic treatment of older persons, deaths from septicemic plague occurred primarily among persons less than 30 years old . Deaths from septicemic plague could be reduced by aggressive antibiotic therapy for patients with a clinical presentation suggesting gram-negative septicemia, especially patients less than 30 years old. Equine Vet J, 1987 Jan, 19(1), 25 - 8 Plasma endotoxin levels in horses subjected to carbohydrate induced laminitis; Sprouse RF et al.; Thirteen (65 per cent) of 20 horses subjected to carbohydrate overload developed Obel Grade 3 lameness within 56 h . Increases in plasma endotoxin from control levels of less than 0.1 ng/litre to values ranging from 2.4 to 81.53 ng/litre were measured in 11 (85 per cent) of 13 horses during the onset of Obel Grade 3 lameness . Obel Grade 3 lameness was associated with rises in plasma Gram-negative endotoxin levels in 11 (92 per cent) of 12 horses . Two peak increases separated by 16 h were verified in five (45 per cent) of 11 horses that exhibited both endotoxaemia and Obel Grade 3 lameness . The first peak occurred, on average, at 32 h and the second peak at 48 h post overload when only one peak was measured, this occurred in four horses at the average time of 24 h whereas in another three horses only a 48 h or 56 h peak was detected after carbohydrate overload. Dev Comp Immunol, 1987 Summer, 11(3), 551 - 64 Bacterial sialic acid modulates activation of the alternative complement pathway of channel catfish (Ictalurus punctatus); Ourth DD et al.; The alternative complement pathway (ACP) provides the non-immune channel catfish (Ictalurus punctatus) with protection against many Gram-negative bacteria . Very little serum bactericidal activity (0-13%) was found against 8 fish pathogens, but a strong bactericidal response (100%) was found against 7 non-pathogens . MgEGTA chelation of catfish serum did not essentially change the bactericidal results . Catfish serum heated at 56 degrees C and serum adsorbed with zymosan had no bactericidal activity . This demonstrated that the ACP was responsible for the bactericidal response . The molecular nature of the microbial surface determines whether or not the ACP will be activated . A relative lack of surface sialic acid has been found to be important for binding complement Factor B of the ACP by susceptible microbial surfaces . This study therefore examined the 15 Gram-negative bacterial fish pathogens and non-pathogens by determining their sialic acid content and their ability to elicit a bactericidal response by the catfish ACP . It was found that there was very little bactericidal activity against the fish pathogens that contained sialic acid but a very strong bactericidal response (100%) against the non-pathogens that lacked sialic acid (p = .0043) . A relative lack of sialic acid or no sialic acid therefore correlated with a strong bactericidal response by the catfish ACP . Neuraminidase treatment of the bacterial fish pathogens to remove sialic acid greatly increased the bactericidal response against them by the catfish ACP when compared with untreated bacteria (p = .0431). Intensive Care Med, 1987, 13(5), 347 - 51 Colonization and infection in surgical intensive care patients--a prospective study; Kerver AJ et al.; Nosocomial infections are a major problem in intensive care patients . Thirty-nine patients, requiring intensive care for 5 days or more (mean 15.8 days) were prospectively investigated, to determine the relation between colonisation and nosocomial infection . Thrice weekly, cultures from the oropharynx, respiratory and digestive tract were obtained . Colonization with aerobic gram-negative microorganisms of the oropharynx, respiratory and digestive tract significantly increased during the stay in the Intensive Care Unit . In 29 patients (74%) 78 nosocomial infections were diagnosed . The most frequent nosocomial infections were pneumonia (26 patients, 66.6%), catheter-related bacteraemia (11 patients, 28.2%), and wound infections (7 patients, 17.9%) . In 59 instances (75.6%), colonization with the same potential pathogenic microorganism preceded the nosocomial infection . The overall mortality was 25.6% (10 patients), bacteraemia with aerobic gram-negative microorganisms being the cause of death in 7 patients. Circ Shock, 1987, 22(4), 333 - 41 Effect of fibronectin supplementation in endotoxic shock in the dog; Hauptman JG et al.; An earlier study by our group demonstrated significant amelioration of hypotension, hypoglycemia, and acidosis in dogs treated with purified human plasma fibronectin prior to induction of endotoxic shock . The present study was completed to determine whether treatment with purified human plasma fibronectin 1 hr after induction of endotoxic shock would provide similar benefits . To this end, selected hemodynamic, pulmonary, acid-base, metabolic, hematological, and serum chemistry parameters were monitored for 6 hours in two groups of anesthetized dogs in Escherichia coli endotoxic shock . One group was given an intravenous injection of purified human plasma fibronectin, and the other received an equal volume of saline 1 hr after shock induction . Between-group analysis of the data revealed no significant differences between any parameter excepting modest differences in plasma glucose, albumin, alkaline phosphatase, and BUN concentrations . However, even these differences, although statistically significant, were sporadic and unimpressive . This study suggested that treatment of dogs with fibronectin during gram-negative endotoxic shock was not efficacious. Circ Shock, 1987, 22(4), 303 - 9 Experimental gram-negative pneumonia produces a hyperdynamic septic profile; Keenan RJ et al.; General anesthesia (GA) and extensive surgery undertaken to reproduce a hyperdynamic septic shock syndrome (HSSS) may confound the observed effects of pure HSSS . A large animal model of HSSS without GA or surgery was created in sheep following production of a Pseudomonas pneumonia by direct bronchoscopic instillation into a dependent lobe using only light ketamine anesthesia . Cardiac output rose significantly (5.05 to 6.32 L/min) while SVR {1,421.4 to 1,000.5 dynes X s X cm(-5)} and mean BP (92.5 to 82.0 mmHg) fell in the septic animals . Systemic infection was confirmed by blood culture . This model reliably produces hyperdynamic sepsis without the confounding effects of GA or extensive surgery. Prog Clin Biol Res, 1987, 231, 405 - 16 Clinical evaluation of the plasma chromogenic Limulus assay; Dolan SA et al.; Clinical correlations of the plasma chromogenic Limulus assay were evaluated in 520 septic episodes to assess the diagnostic utility of the assay in a university hospital setting . Otherwise unselected patients undergoing blood culture were studied . An association of plasma Limulus activity with gram negative bacteremia and focal infections was found (p less than .001 and p less than .01, respectively) . There was a lesser correlation between a positive assay and the presence of major hepato-intestinal illness (p = .05) . The frequency of positive tests was similar for adult and pediatric groups . There were modest correlations of a positive assay with hypotension (p = .02) and thrombocytopenia (p = .04), but only among patients with gram negative infection . Abnormal neutrophil parameters were unassociated with positive assays in any group . The sensitivity and specificity of the test for a condition known to cause endotoxemia--either gram negative infection or major intestinal disease--were low, 21% and 93% respectively . However, the predictive value of a positive test was 79%, indicating utility for the assay. Circ Shock, 1987, 21(1), 1 - 13 Protection against disseminated intravascular coagulation and death by antithrombin-III in the Escherichia coli endotoxemic rat; Emerson TE Jr et al.; Gram-negative septic shock remains a major clinical problem . One frequently encountered complication of sepsis is disseminated intravascular coagulation (DIC) . The present study was to determine in an Escherichia coli endotoxemia awake rat model the efficacy of antithrombin-III (AT-III) prophylaxis and to explore the role of DIC in the pathogenesis of endotoxemia . We demonstrated that DIC occurs very early, before the appearance of detectable serious abnormalities in cardiovascular, metabolic, and biochemical variables indicative of organ damage or dysfunction; AT-III prophylaxis significantly ameliorates DIC, as evidenced by completely preventing the fall in plasma fibrinogen concentration and significantly limiting the increases in prothrombin time and activated partial thromboplastin time after 4 hours of endotoxemia; and AT-III prophylaxis dramatically increases permanent survival . Results of this study suggest that AT-III prophylaxis is very protective above a threshold dosage in an endotoxemic rat model and that protection is in part due to ameliorating DIC . Our data also suggest that DIC occurs very early during endotoxemia and may in part be responsible for the pathogenesis of endotoxemia in the rat . We conclude that AT-III prophylaxis may be efficacious in conditions of impending DIC, such as gram-negative septicemia/endotoxemia. Fed Proc, 1987 Jan, 46(1), 97 - 104 Acute inflammation in gram-negative infection: endotoxin, interleukin 1, tumor necrosis factor, and neutrophils; Movat HZ et al.; Experimental bacterial infection of the dermis induced with gram-negative microorganisms is associated with an acute inflammatory reaction, which represents the principal local defense against spread of the infection . When the inflammatory reaction is quantitated with radiolabeled cells and proteins, the kinetics resemble acute inflammation induced with other agents, such as immune complexes or chemotaxins . There is an interrelationship between the components or events of the inflammatory reaction; inasmuch as vascular injury is neutrophil-dependent, neutrophils must migrate to the site where the bacteria multiply . In neutropenic animals there is no such emigration and bacterial multiplication is not inhibited . The microorganisms shed endotoxin, which in turn induces secretion of interleukin 1 (IL 1) and probably tumor necrosis factor . Endotoxin is the most potent agent (10(-15) mol vs . 10(-12) mol of C5ades Arg) capable of inducing a neutrophil influx . Desensitization or tachyphylaxis of the tissues (probably of postcapillary venular endothelium) to IL 1 seems to control cessation of the neutrophil influx (also in vitro evidence) . Phagocytosis of the bacteria by neutrophils is associated with release of oxygen radicals and lysosomal proteases from the neutrophils . These are instrumental in eliciting microvascular injury, which is characterized by enhanced vasopermeability, hemorrhage, and thrombosis. Lab Invest, 1987 Jan, 56(1), 49 - 59 Role for macrophage products in endotoxin-induced polymorphonuclear leukocyte accumulation during inflammation; Issekutz AC et al.; Endotoxins (lipopolysaccharide, LPS) released by Gram-negative bacteria induce acute inflammation with polymorphonuclear leukocyte (PMNL) infiltration . The mechanism of PMNL accumulation appears to be complement-independent and is not well understood . Here, we report investigation of the factors which may mediate LPS-induced PMNL accumulation in the pleural cavity and skin of rabbits . The intrapleural injection of 50 ng of Escherichia coli 0111 LPS caused the appearance in the exudate fluid of an activity which, upon intradermal injection induced PMNL accumulation in the skin, as measured by a 51Cr-labeled leukocyte assay and which was confirmed histologically . This activity preceded by 30 minutes the massive influx of PMNL into the pleural cavity . 125I-labeled LPS, gel filtration chromatography, limulus amebocyte lysate assays, and polymyxin B allowed distinction between reactions in the skin attributable to LPS and reactions due to the effect of this "PMNL infiltration-inducing activity." Pleural macrophages cultured for 3 to 6 hours with 3 to 30 ng/ml of LPS also released factors which induced PMNL infiltration into the skin . Sephadex G-100 chromatography of LPS-induced pleural exudate fluid or of supernatants from LPS-stimulated macrophage cultures yielded identical elution profiles, with one major peak of PMNL infiltration-inducing activity at an apparent molecular weight of 45,000 and a minor peak at 14,000 to 18,000 . Only the low molecular weight fraction contained interleukin 1 activity . Lipid A was required for the secretion of these factors by macrophages . The LPS shed by killed E . coli also induced macrophage production of PMNL infiltration-inducing activity . The activity was sensitive to pronase, and its production was inhibited by an inhibitor of protein synthesis (cycloheximide) . The active factors did not induce PMNL chemotaxis, aggregation, or chemiluminescence in vitro indicating that the activity was not C5a . We conclude that PMNL infiltration induced by LPS and perhaps by Gram-negative bacteria, may be mediated in part by the secretion from tissue macrophages of factors which can recruit PMNLs from the blood . The most active of these (approximately equal to 45,000 daltons) lacks interleukin-1 or PMNL chemotactic activity. Eur J Clin Pharmacol, 1987, 33(5), 469 - 72 Oral pharmacokinetics and ascitic fluid penetration of pefloxacin in cirrhosis; Cardey J et al.; Plasma and ascitic fluid concentrations of pefloxacin in 10 cirrhotic patients and 8 healthy volunteers were determined following administration of a single oral dose of 400 mg . The mean elimination half-life was significantly increased in the patients (29.0 h) compared to in 8 healthy volunteers (12.3 h) . In patients, the total plasma clearance (2.71 vs 6.85 l/h) and volume of distribution (1.12 vs 1.67 l/kg) were decreased . Estimated by the ratio of the AUC in peritoneal fluid and plasma, ascitic fluid penetration was 68% after one oral dose, and pronounced accumulation of pefloxacin in ascites was found after repeated doses . Oral pefloxacin would seem to be a convenient and useful treatment of spontaneous, gram-negative, bacterial peritonitis in cirrhosis . However, the decreased hepatic metabolism of the drug leads to a marked accumulation in plasma and ascites after repeated doses, and a reduced dose is required in these patients. Med Oncol Tumor Pharmacother, 1987, 4(2), 59 - 66 Iatrogenic and idiopathic acute myelogenous leukemia: a comparison of clinical features and treatment complications; Williams CK et al.; We have compared the clinical and laboratory features as well as treatment complications observed in 6 patients with iatrogenic acute myelogenous leukemia (I-AML) with those of 26 patients with idiopathic acute myelogenous leukemia (AML) . I-AML patients were significantly younger and their disease appeared less virulent on admission than in the AML patients . Following identical chemotherapy,